JP7619555B2 - Administration kit - Google Patents

Description

This disclosure relates to an administration kit.

Cell transplantation therapy, in which cells are administered into the body for the treatment of humans and animals, is known, and studies are also being conducted on administration kits for administering cells into the body. For example, Patent Document 1 describes a cell injection device equivalent to an administration kit. The cell injection device of Patent Document 1 is a cell injection device for injecting (equivalent to administration) cells into the heart for cell therapy of heart disease, and discloses a configuration including an injection needle (equivalent to a puncture needle) that is inserted into biological tissue, a tube equipped with the injection needle at its tip, and a syringe connection part provided on the base end side of the tube. According to the cell injection device of Patent Document 1, a cell syringe containing cells is connected to the syringe connection part, and the cells are pushed into the tube by the cell syringe, thereby injecting the cells into the biological tissue through the tube.

JP 2011-067588 A

In cell transplantation therapy, in addition to cells, cell structures may be administered into the body. Here, the cell structure is a structure in which multiple cells and multiple biocompatible polymer blocks are mixed, and has a structure in which multiple polymer blocks are arranged in the gaps between multiple cells. The cell structure is larger in size than cells. When cells or cell structures are administered into the body, they are administered in the form of a suspension in which the cells or cell structures are suspended in a liquid such as artificial cerebrospinal fluid (hereinafter referred to as a cell suspension).

In such a procedure for administering a cell suspension into a living body, it can take a very long time from the insertion of the puncture needle to the administration of the entire amount of the cell suspension to be administered. For example, when administering a cell suspension to the brain, not only must the needle be inserted carefully, but the cell suspension must also be administered in small amounts at time intervals. In this case, it can take a considerable amount of time from the start of administration of the cell suspension to the completion of administration of the entire amount of the cell suspension. This can lead to the following problems:

That is, when performing the procedure of administering a cell suspension, first, as a preparatory step, a cell suspension is prepared by dispersing cells or cell structures in a liquid such as artificial cerebrospinal fluid. The prepared cell suspension is then injected into a syringe, and the cell suspension is contained in the syringe. In this state, a puncture needle is connected to the syringe. After completing these preparatory steps, the procedure of administering the cell suspension begins.

As described above, the procedure for administering a cell suspension begins with inserting the puncture needle into the living body, and administration of the cell suspension into the living body begins after the puncture needle has been inserted. In this way, after the cell suspension is contained in the syringe, it may take more than five minutes before the cell suspension in the syringe is actually administered to the living body, because of the steps of connecting the puncture needle to the syringe and inserting the puncture needle into the living body.

If the cell suspension is held in the syringe for a long time, the cells or cell structures dispersed in the cell suspension may settle within the syringe. If the cells or cell structures settle within the syringe, there is a risk that the cells or cell structures may become clogged at the tip of the syringe or at the puncture needle, or at other parts of the syringe that have a smaller diameter than the inner diameter of the syringe body. If the cells or cell structures become clogged, the cell suspension cannot be administered smoothly.

Cited Reference 1 does not disclose or suggest these issues or their solutions.

The present disclosure has been made in consideration of the above circumstances, and aims to provide an administration kit that can smoothly administer a cell suspension to a living body by reducing clogging caused by precipitation of cells or cell structures more than ever before.

The administration kit of the present disclosure includes the following aspects.
＜1＞
The present invention relates to an administration tube used for administering a cell suspension containing cells or cell structures into a living body, the administration tube having a first end to which a puncture needle can be attached and a second end to which a syringe can be attached, and including an administration tube capable of holding therein the entire amount of the cell suspension to be administered in one treatment, and the entire amount of the cell suspension to be administered in one treatment.
＜2＞
The administration kit according to <1>, comprising a container for containing a cell suspension to be filled into an administration tube.
＜3＞
An administration kit according to <1> or <2>, comprising a cell suspension supply syringe that can be attached to the second end of the administration tube and is used to supply the cell suspension into the administration tube.
＜4＞
An administration kit as described in <1>, comprising a cell suspension supply syringe that can be attached to the second end of the administration tube and is used to supply the cell suspension into the administration tube, and the cell suspension supply syringe is filled with the entire amount of the cell suspension.
＜5＞
The administration kit according to <1>, wherein the entire amount of the cell suspension is filled in an administration tube.
＜6＞
An administration kit described in any one of <1> to <4>, including an instruction manual for instructing a user to start administration of the cell suspension after filling the administration tube with the entire amount of the cell suspension to be administered in one treatment.
＜７＞
The instruction manual includes a first instruction to rotate the cell suspension supply syringe back and forth multiple times around its central axis before starting to supply the cell suspension from the cell suspension supply syringe filled with the entire amount of cell suspension to be administered in one treatment to the administration tube, and a second instruction to fill the entire amount of the cell suspension into the administration tube by making the cell suspension in the cell suspension supply syringe uniformly dispersed and then starting to supply the cell suspension from the cell suspension supply syringe to the administration tube.
＜8＞
The administration kit described in <7>, wherein the instruction manual includes a third instruction to supply the entire amount of the cell suspension to the administration tube within 2 seconds after the cell suspension is uniformly dispersed.
＜9＞
The administration kit according to any one of <1> to <8>, wherein the administration tube has a capacity of 2 mL or more.
＜10＞
The administration kit according to any one of <1> to <9>, wherein the administration tube has an inner diameter of 1.0 mm or more and 1.7 mm or less.
<11>
The administration kit according to any one of <1> to <10>, wherein the administration tube has a length of 3 m or less.
＜12＞
The administration kit according to any one of <1> to <11>, wherein the administration tube has a capacity capable of accommodating the cell suspension that is greater than the total volume of the cell suspension to be administered in one treatment.
＜13＞
The administration kit according to any one of <1> to <12>, wherein the administration tube has a puncture needle attachment portion for attaching a puncture needle to a first end thereof.
＜14＞
The administration kit according to any one of <1> to <13>, wherein the administration tube has a syringe attachment portion for attaching a syringe to a second end thereof.
＜15＞
The syringe mounting portion includes a plurality of mounting ports to which syringes can be attached, and a switching valve disposed between the administration tube and the mounting ports to switch a communication state between the administration tube and the plurality of mounting ports.
＜16＞
The administration kit according to any one of <1> to <15>, wherein the cell suspension contains a cell structure, and the cell structure has a diameter of 100 μm or more.
＜１７＞
The administration kit according to <16>, wherein the diameter of the cell structure is 100 μm or more and 500 μm or less.
＜18＞
The administration kit according to <16> or <17>, wherein the cell structure comprises a biocompatible polymer block and at least one type of cell, and a plurality of polymer blocks are arranged in the gaps between a plurality of cells.
＜19＞
The administration kit according to any one of <1> to <18>, further comprising an extrusion liquid supply syringe that can be attached to the second end of the administration tube and that holds an extrusion liquid for extruding the cell suspension filled in the administration tube.
＜20＞
The administration kit according to <19>, wherein the extrusion force for extruding the extrudate from the extrudate supply syringe at an extrusion rate of 12 mL/min is 15 N or less.
＜21＞
The administration kit according to any one of <1> to <20>, further comprising a puncture needle attached to the first end of the administration tube.
＜22＞
The puncture needle includes a needle body along a front-rear direction, with a passage for a cell suspension formed therein, and an outlet for the cell suspension formed on an outer surface of a front end of the needle body,
the puncture needle includes a connection portion connected to a rear end of the needle body and an administration tube, the connection portion including a body insertion hole into which the rear end of the needle body is inserted, and a tube insertion hole communicating with the body insertion hole and into which a first end of the administration tube is inserted;
The administration kit described in <21>, wherein the tip side portion of the tube insertion hole connected to the main body insertion hole has a funnel shape whose inner diameter gradually narrows from the base end side to the tip side, and the opening angle of the funnel shape opening from the tip side to the base end side is 120° or less.
＜23＞
The administration kit according to <22>, wherein the puncture needle has an inner diameter of a passage of the needle body of 0.5 mm or more.
＜24＞
The administration kit according to any one of <22> and <23>, wherein the inner diameter of the passage of the needle body of the puncture needle is at least twice the diameter of the cells or cell structures contained in the cell suspension.
＜25＞
The administration kit according to any one of <22> to <24>, wherein the opening area of the outlet of the puncture needle is equal to or larger than the cross-sectional area of the passage of the needle body.

In the administration kit of the present disclosure, the puncture needle includes a needle body along the front-rear direction, with a passage for a cell suspension formed therein, and an outlet for the cell suspension formed on the outer surface of the front end of the needle body,
the needle body is a single tube with a passageway;
The cross-sectional area of the outlet passage perpendicular to the direction in which the outlet passage extends may smoothly increase toward the outlet port in at least a portion of the outlet passage, and may be greater than or equal to the cross-sectional area at each position in the direction in which the outlet passage extends than at a position closer to the passage.

In the administration kit of the present disclosure, the puncture needle is preferably made of a metal such as stainless steel or aluminum, ceramic, or an inflexible hard resin.

In the administration kit of the present disclosure, the puncture needle has a connection part at the rear end of the needle body that is connected to the administration tube, the distance between the tip of the connection part and the tip of the needle body part is 170 mm to 220 mm, and the needle body may have an outer diameter of 1.6 mm or less.

In the administration kit of the present disclosure, the outer surface of the front end of the puncture needle may include a front end face that protrudes forward while smoothly curving, and a cylindrical side face connected to the front end face.

In the administration kit of the present disclosure, the front end of the outlet of the puncture needle may be located rearward of the front end of the passage, and may be located at the boundary between the front end face and the side face or rearward of the boundary.

In the administration kit of the present disclosure, the puncture needle may have an outlet formed behind the front end surface.

In the administration kit of the present disclosure, the puncture needle has a cylindrical side surface at the front end,
At least a part of the outlet may be formed on the side surface.

In the administration kit of the present disclosure, the outer surface of the needle body of the puncture needle is the surface that comes into contact with the living body when the needle is inserted, and it is preferable that the wall portion from the passage of the needle body to the outer surface is made of a single layer.

In the administration kit of the present disclosure, the puncture needle has an outlet path that includes a cylindrical portion that connects to the passage and a truncated cone portion that connects the outlet and the cylindrical portion, the inner diameter of the passage is 0.3 mm to 1.2 mm, the inner diameter of the cylindrical portion is 0.3 mm to 1.2 mm, and the cone angle of the truncated cone portion may be 90°.

In the administration kit of the present disclosure, the puncture needle may have an outlet path that further includes a second cylindrical portion between the outlet and the truncated cone portion, and the inner diameter of the outlet and the maximum inner diameter of the truncated cone portion may be 1.2 mm to 1.6 mm.

In the administration kit of the present disclosure, the puncture needle has an outlet path that includes, between the passage and the outlet, a truncated cone portion or a cylindrical portion of a first size, and a truncated cone portion or a cylindrical portion of a second size larger than the first size, from the passage side toward the outlet, and the first size may be 0.3 mm to 1.2 mm, and the second size may be 1.2 mm to 1.66 mm.

In the administration kit of the present disclosure, the puncture needle may have an outlet path that includes a truncated cone portion that is connected to the passage and a cylindrical portion that is provided between the outlet and the truncated cone portion, and the inner diameter of the cylindrical portion may be 0.3 mm to 1.2 mm, and the maximum inner diameter and outlet of the truncated cone portion may be 1.2 mm to 1.6 mm.

In the administration kit of the present disclosure, the puncture needle is inclined at an angle θ greater than 0° and in the range of 30° so that the outlet side is located further forward than the passage side, and the inner diameter of the passage side may be 0.3 mm to 1.2 mm, and the size of the outlet may be 0.5 mm to 1.2 mm.

In the administration kit of the present disclosure, it is preferable that the puncture needle suppresses the generation of a jet of water when the water is injected into the passage from the syringe up to a water injection pressure of 5 N.

In the administration kit of the present disclosure, the entire amount of the cell suspension may be filled in the administration tube for 5 minutes or more.

In the administration kit of the present disclosure, the entire amount of the cell suspension may be filled in the administration tube for 10 minutes or more.

The administration kit of the present disclosure may be an administration kit for head injection.

The administration kit of the present disclosure may be an administration kit for brain treatment.

The administration kit disclosed herein reduces clogging caused by precipitation of cells or cell structures, making it possible to administer cell suspensions to living organisms more smoothly than ever before.

FIG. 1 is a schematic diagram of an administration kit according to one embodiment. FIG. 1 is a first explanatory diagram of a method of using the administration kit. FIG. 2 is a second explanatory diagram of a method of using the administration kit. FIG. 3 is a third explanatory diagram of a method for using the administration kit. FIG. 4 is a fourth explanatory diagram of a method for using the administration kit. FIG. 1 is an explanatory diagram of the presence or absence of air bubbles in a dispensing tube and clogging of cell structures. FIG. 1 is a schematic diagram of an administration kit according to a modified example. FIG. 2 is a front view of an example puncture needle. FIG. 2 is a partial enlarged view of the puncture needle of FIG. 1 including a partial cross section of the puncture needle. Fig. 10A is a partially enlarged view of Fig. 9. Fig. 10B is a plan view of Fig. 10A. FIG. 11 is a cross-sectional view of a puncture needle according to a modified example of the design. Fig. 12A is a plan view of the puncture needle 1A, Fig. 12B is a cross-sectional view taken along line BB in Fig. 12A, and Fig. 12C is an enlarged view of the dashed line region in Fig. 12B. 13 is a photograph showing the change over time of the cell suspension in a cell suspension supply syringe. FIG. 1 is an explanatory diagram of an administration kit according to Example 2. FIG. 13 is an explanatory diagram of the administration kit of Example 3. FIG. 1 is an explanatory diagram of an administration kit according to Example 4. FIG. 13 is an explanatory diagram of the administration kit of Example 5. Fig. 18A is a plan view of the puncture needle 1B, Fig. 18B is a cross-sectional view taken along line BB in Fig. 18A, and Fig. 18C is an enlarged view of the dashed line region in Fig. 18B. Fig. 19A is a plan view of the puncture needle 1C, Fig. 19B is a cross-sectional view taken along line BB in Fig. 19A, and Fig. 19C is an enlarged view of the dashed line region in Fig. 19B.

Hereinafter, the embodiments of the present disclosure will be described in detail with reference to the drawings. In this disclosure, "about" means a value including a range of 10% above and below.

"Administration kit"
FIG. 1 is a schematic diagram showing an administration kit 5 of one embodiment. The administration kit 5 of this embodiment includes a puncture needle 1, an administration tube 2, and an amount of cell suspension 3 to be administered in one treatment. For example, the administration kit 5 is manufactured by a manufacturer and provided to medical facilities such as hospitals as medical consumables used in treatment. The administration kit 5 is used for cell transplantation treatment. As an example, the administration kit 5 is used to perform cell transplantation into a specific part of the brain of a living body for the treatment of cerebral infarction, etc. The administration kit 5 may be for head injection or for brain treatment.

The puncture needle 1 is a puncture needle for injecting the cell suspension 3 into the central nervous system. When performing cell transplantation into the brain, the puncture needle 1 is attached to a stereotactic brain surgery device 100 (see Figure 5) and used. Details of the puncture needle 1 will be described later.

The administration tube 2 is a tube used for administering a cell suspension containing cells or cell structures 3a into a living body. The administration tube 2 is flexible. The administration tube 2 has a first end 2a to which the puncture needle 1 can be attached, and a second end 2b to which a syringe can be attached. The administration tube 2 has a capacity capable of holding the entire amount of cell suspension 3 to be administered in one treatment. The administration tube 2 has an inner diameter smaller than the inner diameter of a cell suspension supply syringe 6 (described below) used to supply the cell suspension 3 to the administration tube 2.

The administration tube 2 preferably has an inner diameter of 1.0 mm or more and 1.7 mm or less. The administration tube 2 only needs to have a capacity at least large enough to hold the entire amount of the cell suspension 3 to be administered in one treatment. However, it is more preferable that the capacity is larger than the volume of the entire amount of the cell suspension to be administered in one treatment. The capacity of the administration tube 2 can be, for example, 2 mL or more. It is preferable that the administration tube 2 has a length of 3 m or less.

In this example, the administration tube 2 has a coiled body as shown in FIG. 1. A tube wound in a coil shape like this is also called a spiral tube. If the administration tube 2 has a coiled body, it is easy to handle because the overall length can be kept compact even if it is made long to increase the capacity. For this reason, it is preferable that the administration tube 2 has a coiled spiral tube body as in this example. Although it is preferable that the administration tube 2 is a spiral tube like this, it does not necessarily have to be a spiral tube, and it may be a general medical infusion tube that is straight and not coiled.

The first end 2a of the administration tube 2 is provided with a puncture needle attachment part 26 for attaching the puncture needle 1, and the puncture needle 1 is attached to the first end 2a via the puncture needle attachment part 26. The second end 2b of the administration tube 2 is provided with a syringe attachment part 28 for attaching a syringe, and the syringe is attached to the second end 2b via the syringe attachment part 28.

The cell suspension 3 is a suspension in which cells or cell structures 3a are suspended in a liquid such as artificial cerebrospinal fluid. In the cell suspension 3, the cells or cell structures 3a are dispersed in a state where they are not dissolved in the liquid. Details of the cell suspension 3 will be described later. In this example, the cell suspension 3 will be described as containing cell structures 3a. The cell suspension 3 to be filled into the administration tube 2 is contained in a container 4. That is, the container 4 is, for example, a vial, and contains the amount of cell suspension 3 to be administered in one treatment.

Here, the "total amount of cell suspension 3 to be administered in one treatment" refers to the amount of cell suspension 3 determined according to the therapeutic objective of the cell transplantation treatment, and refers to the total amount of cell suspension 3 to be administered from the time the puncture needle 1 is inserted into the living body until the puncture needle 1 is removed from the living body. The administration tube 2 may be filled with the total amount of cell suspension 3 for 5 minutes or more, or may be filled with the total amount of cell suspension 3 for 10 minutes or more.

"Cell Structure"
The cell structure 3a is a structure in which a plurality of biocompatible polymer blocks are arranged in the gaps between a plurality of cells and a biocompatible polymer block. Here, biocompatible means that when it comes into contact with a living body, it does not cause significant adverse reactions such as long-term and chronic inflammatory reactions. The biocompatible polymer used in the technology of the present disclosure is not particularly limited as to whether it is decomposed in the living body as long as it has affinity for the living body, but it is preferably a biodegradable polymer. For example, the one described in International Publication No. 2017/221879 can be used as the cell structure 3a.

(Biocompatible polymer block)
The biocompatible molecular block is a mass made of a biocompatible polymer.

The shape of the biocompatible polymer block is not particularly limited. For example, it may be amorphous, spherical, particulate (granular), powdery, porous, fibrous, spindle-shaped, flat, or sheet-shaped, and preferably amorphous, spherical, particulate (granular), powdery, or porous. The term "irregular" refers to a surface shape that is not uniform, for example, an object with irregularities like a rock. Note that the above examples of shapes are not separate, and for example, the term "irregular" may be used as an example of a sub-concept of particulate (granular). It is particularly preferable that the biocompatible polymer block is in the form of granules obtained by crushing a porous body of biocompatible polymer.

The size of each biocompatible polymer block is preferably 20 μm or more and 200 μm or less, and more preferably 50 μm or more and 120 μm or less.

The biocompatible polymer may be crosslinked or not, but is preferably crosslinked. The use of a crosslinked biocompatible polymer has the effect of preventing instantaneous decomposition during culture in a medium and transplantation into a living body. The crosslinking method includes, but is not limited to, thermal crosslinking, crosslinking with aldehydes (e.g., formaldehyde, glutaraldehyde, etc.), crosslinking with condensing agents (carbodiimide, cyanamide, etc.), enzymatic crosslinking, photocrosslinking, ultraviolet crosslinking, hydrophobic interactions, hydrogen bonds, and ionic interactions. Thermal crosslinking, ultraviolet crosslinking, or enzymatic crosslinking are preferred.

The biocompatible polymer is preferably gelatin, collagen, atelocollagen, elastin, fibronectin, pronectin, laminin, tenascin, fibrin, fibroin, entactin, thrombospondin, retronectin (registered trademark), polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, hyaluronic acid, glycosaminoglycan, proteoglycan, chondroitin, cellulose, agarose, carboxymethylcellulose, chitin, or chitosan.

The biocompatible polymer is particularly preferably recombinant gelatin. Recombinant gelatin refers to a polypeptide or protein-like substance having an amino acid sequence similar to that of gelatin, produced by genetic recombination technology. The following forms of recombinant gelatin are preferred:

The recombinant gelatin is preferably represented by the following formula 1:
Formula 1: A-[(Gly-X-Y)n]m-B
In the formula, A represents any amino acid or amino acid sequence, B represents any amino acid or amino acid sequence, n Xs each independently represent any amino acid, n Ys each independently represent any amino acid, n represents an integer from 3 to 100, and m represents an integer from 2 to 10. The n Gly-X-Ys may be the same or different.

Recombinant gelatin is
(1) a peptide consisting of the amino acid sequence set forth in SEQ ID NO:1;
(2) a peptide having biocompatibility and consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence set forth in SEQ ID NO:1; or (3) a peptide having biocompatibility and consisting of an amino acid sequence having 80% or more sequence identity with the amino acid sequence set forth in SEQ ID NO:1;
It is particularly preferred that:

It is preferable that the cell structure 3a contains 0.0000001 μg or more and 1 μg or less of a biocompatible polymer block per cell.

The thickness or diameter of the cell structure 3a is preferably 50 μm or more, and preferably 1 mm or less. The thickness or diameter of the cell structure 3a is preferably in the range of 80 μm or more and 800 μm or less, more preferably 100 μm or more and 500 μm or less. In this specification, the "thickness or diameter of the cell structure 3a" refers to the following. When a certain point P in the cell structure 3a is selected, the length of the line segment that divides the cell structure 3a so that the distance from the outer shell of the cell structure 3a is the shortest among the straight lines passing through the point P is defined as line segment A. A point P in the cell structure 3a where the line segment A is the longest is selected, and the length of line segment A at that time is defined as the "thickness or diameter of the cell structure". In other words, the "thickness or diameter of the cell structure" refers to the maximum diameter of the cell structure 3a.

(cell)
The type of cells is not particularly limited, and any cell can be used depending on the actual purpose of treatment. The cells used may be one type or a combination of multiple types of cells. The cells used are preferably animal cells, more preferably vertebrate-derived cells, and particularly preferably human-derived cells. The type of vertebrate-derived cells (particularly human-derived cells) may be any of pluripotent cells, somatic stem cells, progenitor cells, or mature cells, and particularly preferably somatic stem cells.
As the somatic stem cells, for example, mesenchymal stem cells (MSCs), hematopoietic stem cells, amniotic cells, umbilical cord blood cells, bone marrow-derived cells (e.g., bone marrow-derived mesenchymal stem cells, etc.), cardiac stem cells, adipose-derived stem cells, or neural stem cells can be used. As the precursor cells and mature cells, for example, cells derived from the skin, dermis, epidermis, muscle, cardiac muscle, nerve, bone, cartilage, endothelium, brain, epithelium, heart, kidney, liver, pancreas, spleen, oral cavity, cornea, bone marrow, umbilical cord blood, amniotic membrane, or hair can be used. As the human-derived cells, for example, ES cells, iPS cells, MSCs, chondrocytes, osteoblasts, osteoblast precursor cells, mesenchymal cells, myoblasts, cardiac muscle cells, cardiac muscle blasts, nerve cells, hepatocytes, beta cells, fibroblasts, corneal endothelial cells, vascular endothelial cells, corneal epithelial cells, amniotic cells, umbilical cord blood cells, bone marrow-derived cells, or hematopoietic stem cells can be used. In addition, the origin of the cells may be either autologous or allogeneic cells. Among the above, it is preferable to use mesenchymal stem cells as the cells. As the mesenchymal stem cells, adipose-derived mesenchymal stem cells or bone marrow-derived mesenchymal stem cells are preferable. As the origin of the mesenchymal stem cells, it is preferable to use human-derived or canine-derived.

"How to use the administration kit"
An example of a method of using the administration kit 5 will be described with reference to Figures 2 to 5. In addition to the administration kit 5, a cell suspension supply syringe 6 and an initial filling liquid and extrusion liquid supply syringe 7 are prepared (see Figures 2 and 3). The initial filling liquid and extrusion liquid supply syringe 7 is used to supply the initial filling liquid and extrusion liquid.

The initial filling liquid is filled into the puncture needle 1 and the administration tube 2 before the cell suspension 3 is supplied to the administration tube 2 so that there are no air bubbles in the puncture needle 1 and the administration tube 2 when the cell suspension is supplied.

The extrusion liquid is used to extrude the cell suspension 3 filled in the administration tube 2 from the puncture needle 1.

The initial filling liquid and the extrusion liquid are liquids that are harmless when administered to a living body, such as physiological saline and surgical washing and perfusion liquid (artificial cerebrospinal fluid). The same liquid can be used as the initial filling liquid and the extrusion liquid. In addition, the initial filling liquid and the extrusion liquid can be the same liquid used for suspending the cell suspension 3. Here, the same liquid is used as the initial filling liquid and the extrusion liquid. For convenience, the initial filling liquid and the extrusion liquid are collectively referred to as artificial cerebrospinal fluid 8. In this example, the initial filling liquid and the extrusion liquid supply syringe 7 is filled with artificial cerebrospinal fluid 8.

The cell suspension 3 in the container 4 is filled into the cell suspension supply syringe 6. In other words, the entire amount of cell suspension 3 to be administered in one treatment is filled into the cell suspension supply syringe 6.

First, as shown in FIG. 2, the puncture needle 1 is attached to the first end 2a of the administration tube 2 via the puncture needle attachment part 26, and the initial filling liquid and extrusion liquid supply syringe 7 filled with artificial cerebrospinal fluid 8 as an initial filling liquid is attached to the second end 2b via the syringe attachment part 28. By supplying the artificial cerebrospinal fluid 8 from the initial filling liquid and extrusion liquid supply syringe 7 to the administration tube 2, the puncture needle 1 and administration tube 2 are filled with the artificial cerebrospinal fluid 8 and are free of air bubbles.

Then, remove the initial filling liquid and extrusion liquid supply syringe 7 from the syringe attachment part 28, and attach the cell suspension supply syringe 6 while taking care not to introduce air bubbles (see Figure 3).

Next, as shown in FIG. 3, the cell suspension supply syringe 6 is repeatedly rotated back and forth around the central axis. By repeating this back and forth rotation, the cell structures 3a in the cell suspension 3 in the cell suspension supply syringe 6 are uniformly dispersed, and then the entire amount in the cell suspension supply syringe 6 is supplied to the administration tube 2 within a short period of time (FIG. 4). The short period of time is preferably 2 seconds or less, and more preferably 1 second or less. As a result, the administration tube 2 is filled with the entire amount of the cell suspension 3 to be administered in one treatment. It is preferable to perform the back and forth rotation multiple times to disperse the cell structures 3a. The number of back and forth rotations is preferably 5 or more, and more preferably 10 or more. The uniformly dispersed state refers to a state in which the cells and cell structures are not precipitated in the cell suspension and are mixed with the liquid.

Then, the cell suspension supply syringe 6 is removed from the administration tube 2, and as shown in FIG. 5, the initial filling liquid and extrusion liquid supply syringe 7 is attached to the second end 2b of the administration tube 2 via the syringe attachment portion 28. With the initial filling liquid and extrusion liquid supply syringe 7 attached to the second end 2b of the administration tube 2, the puncture needle 1 is inserted into the site of the living body where the cell suspension is to be injected.

More specifically, as shown in FIG. 5, the cell suspension 3 is administered to the brain of a living body 102 with the puncture needle 1 attached to the stereotactic brain surgery device 100. For example, when transplanting cells such as stem cells to a specific location in the brain, the stereotactic brain surgery device 100 is attached to the head of a patient whose skull has been perforated by burr hole surgery. The stereotactic brain surgery device 100 holds the insertion position and direction of the puncture needle 1 relative to the brain at a specific position and direction. The puncture needle 1 is attached to a movable part of the stereotactic brain surgery device 100, and the insertion position and direction of the puncture needle 1 are adjusted while held by the movable part.

When inserting the puncture needle 1 into the brain, the puncture needle 1 is inserted into the brain from the brain surface through the perforation while being held by the stereotactic brain surgery device, and the tip of the puncture needle 1 is allowed to reach the desired location in the brain. Note that since puncturing the brain must be done carefully, it generally takes about 10 minutes for the tip to reach the desired location in the brain. After the administration tube 2 is filled with the cell suspension 3, the administration tube 2 is left stationary until the puncture needle 1 is inserted into the living body and its tip reaches the desired location.

After the tip of the puncture needle 1 reaches the target position, the initial filling liquid and extrusion liquid supply syringe 7 is operated to supply artificial cerebrospinal fluid 8 as an extrusion liquid to the administration tube 2. The artificial cerebrospinal fluid 8 is supplied to the administration tube 2 until all of the cell suspension 3 in the administration tube 2 is replaced with the artificial cerebrospinal fluid 8. The cell suspension 3 in the administration tube 2 is pushed out by the artificial cerebrospinal fluid 8 supplied from the initial filling liquid and extrusion liquid supply syringe 7, and is administered into the living body.

The administration kit 5 of the present disclosure includes an administration tube 2 capable of holding the entire amount of cell suspension 3 to be administered in one treatment, and can fill the administration tube 2 with the entire amount of cell suspension 3 to be administered in one treatment. The administration tube 2 has a smaller inner diameter than the cell suspension supply syringe 6, and even if the cell structures 3a of the cell suspension 3 in the administration tube 2 sink, the cell structures 3a can maintain a substantially uniform distribution in the length direction. When the cell suspension is supplied directly to the puncture needle 1 from the cell suspension supply syringe 6 without the administration tube 2 as in the past, or when the administration tube 2 is too long to hold the entire amount of the cell suspension 3, at least a portion of the cell suspension 3 is held in the cell suspension supply syringe 6 that is left stationary for a certain period of time for puncture.

As will be described in detail in the examples below, when the cell suspension supply syringe 6 is moved to uniformly disperse the cell structures 3a inside it, and then the cell suspension supply syringe 6 is stopped, the cell structures 3a begin to sink immediately after the cell suspension supply syringe 6 is stopped. When the cell structures 3a are uniformly dispersed in the cell suspension 3, the transparency of the cell suspension 3 is uniform in the cell suspension supply syringe 6. However, after about 5 seconds have passed after the cell suspension supply syringe 6 is stopped, the transparency becomes higher in the vertical upper part of the cell suspension 3, indicating that the cell structures 3a are in a sparse state, while the transparency becomes lower in the vertical lower part, indicating that the cell structures 3a are in a dense state. This change in transparency progresses to a degree that can be visually confirmed. In other words, after about 5 seconds, the progress of the sinking of the cell structures 3a reaches a degree that can be visually confirmed. After about 10 seconds, the transparency in the vertical direction in the cell suspension 3 becomes even higher, and the transparency in the vertical direction in the downward direction becomes even lower. This indicates that more cell structures 3a have sunk. After about 20 seconds, the cell structures 3a have sunk completely, and are now settled in the vertically lower part of the cell suspension supply syringe 6 (see FIG. 13).

In this state where the cell structures 3a have precipitated, the cell structures 3a clog the outlet of the cell suspension supply syringe 6, causing a problem that the cell structures 3a are not supplied to the puncture needle 1 or the administration tube 2. In contrast, the administration kit 5 includes an administration tube 2 capable of holding the entire amount of cell suspension to be administered in one treatment inside. This allows the entire amount of cell structures 3a to be administered in one treatment to be supplied to the administration tube 2 before the cell structures 3a precipitate in the cell suspension supply syringe 6. Therefore, since there is no need to hold the cell suspension 3 in the cell suspension supply syringe 6 during puncture, it is possible to suppress the occurrence of problems such as the cell structures 3a clogging the outlet of the cell suspension supply syringe 6 or being left in the cell suspension supply syringe 6. In addition, since the inner diameter of the administration tube 2 is smaller than the inner diameter of the cell suspension supply syringe 6, the cell structures 3a can be maintained in a substantially uniformly distributed state in the length direction even if the administration tube 2 is left stationary after the cell structures 3a are filled in a substantially uniformly distributed state in the length direction. Therefore, clogging of the cell structures 3a can be suppressed even when the puncture needle 1 is injected from the administration tube 2. As a result, the administration kit 5 of this example reduces clogging due to precipitation of the cell structures 3a compared to conventional methods, making it possible to administer the cell suspension 3 to the living body 102 smoothly. In this example, the cell structures 3a are used, but cells may be used instead of the cell structures 3a. However, since the particle size of the cell structures 3a is larger than that of cells, the cell structures 3a are more likely to clog. Therefore, the administration kit 5 of this example is particularly effective when using the cell structures 3a.

If the inner diameter of the administration tube 2 is 1.0 mm or more, it is possible for cell structures 3a of a certain size, for example, with a diameter of about 300 μm or less, to pass through without clogging. If it is 1.7 mm or less, the number of cell structures 3a overlapping at the same length position within the administration tube 2 is limited, so that the distribution of the cell structures 3a in the length direction can be made more uniform, and a state of uniform distribution in the length direction within the administration tube 2 can be maintained.

Although the administration kit 5 in the above embodiment includes the puncture needle 1, the administration kit 5 may not include the puncture needle 1, and the puncture needle 1 may be prepared separately, similar to the cell suspension supply syringe 6 and the initial filling liquid and extrusion liquid supply syringe 7. In other words, the administration kit 5 only needs to include at least the administration tube 2 and the amount of cell suspension 3 to be administered in one treatment.

The administration kit 5 of the above embodiment includes a vial as the container 4 for containing the cell suspension 3 to be filled into the administration tube 2, but the container 4 may be an ampoule. Alternatively, the cell suspension may be contained within the tube. By containing the cell suspension 3 in the container 4, handling during storage, transportation, etc. is easy.

In the method of using the administration kit 5 in the above embodiment, a form has been described in which the cell suspension supply syringe 6 and the initial filling liquid and extrusion liquid supply syringe 7 are prepared separately from the administration kit 5, but the administration kit may include one or both of the cell suspension supply syringe 6 and the initial filling liquid and extrusion liquid supply syringe 7.

The cell suspension 3 is not limited to being contained in the container 4, and may be contained in the administration kit 5 in a state of being contained in the cell suspension supply syringe 6. That is, the administration kit 5 may include the cell suspension supply syringe 6 containing the cell suspension 3 instead of the container 4 containing the cell suspension 3. Since there is no need to fill the cell suspension 3 from the container 4 into the cell suspension supply syringe 6, it is possible to prevent the introduction of foreign matter and to save time and effort.

Furthermore, the cell suspension 3 may be contained in the administration tube 2 and included in the administration kit 5. That is, the administration kit 5 may not include the container 4 containing the cell suspension 3, but may include the administration tube 2 filled with the cell suspension 3. Since it is not necessary to fill the administration tube 2 with the cell suspension 3 using the cell suspension supply syringe 6, processing such as uniformly dispersing the cell structures 3a in the cell suspension 3 in the cell suspension supply syringe 6 is not required, and the time until puncture begins can be shortened.

In addition, the inner diameter of the initial filling liquid and extrusion liquid supply syringe 7, the inner diameter of the administration tube 2, and the inner diameter of the puncture needle 1 are preferably 15 N or less, more preferably 10 N or less, and even more preferably 5 N or less, in order to extrude the extrusion liquid at an extrusion rate of 12 mL/min when the initial filling liquid and extrusion liquid supply syringe 7, the administration tube 2, and the puncture needle 1 are filled with water at 25°C. For example, when the average diameter of the cell structure 3a is 250 μm, the above extrusion force condition can be satisfied by setting the inner diameter of the initial filling liquid and extrusion liquid supply syringe 7 to 15.8 mm, the inner diameter of the administration tube 2 to 1.5 mm, and the inner diameter of the puncture needle 1 to 1.1 mm. The extrusion force is defined as a value measured by pushing the syringe through a push-pull gauge.

In the method of use described above, the administration tube 2 is filled with artificial cerebrospinal fluid 8 so that there are no air bubbles in the administration tube 2, and air bubbles are prevented from entering when changing the syringe for the following reasons.

If there are air bubbles in the administration tube 2, the cell structures 3a supplied to the administration tube 2 will not be uniformly dispersed in the longitudinal direction within the administration tube 2, and the cell structures 3a will gather in the area adjacent to the air bubbles upstream of the air bubbles. Figure 6 is a conceptual diagram showing the state of the tip position of the artificial cerebrospinal fluid 8 after the artificial cerebrospinal fluid is supplied when there are no air bubbles in the administration tube 2 (Figure 6A) and when there are air bubbles (Figure 6B). As shown in Figure 6B, if the cell structures 3a reach the entrance to the puncture needle 1 in a state of high concentration adjacent to the air bubbles, they may become clogged at the entrance to the puncture needle 1. To avoid this, it is preferable to make the administration tube 2 free of air bubbles before supplying the cell suspension 3 to the administration tube 2, and to prevent air bubbles from entering when replacing the initial filling liquid and extrusion liquid supply syringe 7 with the cell suspension supply syringe 6.

FIG. 7 is a schematic diagram of an administration kit according to a modified example.
The administration kit 5A shown in Fig. 7 includes a puncture needle 1, an administration tube 2, a container 4 containing an amount of cell suspension 3 to be administered in one treatment, a cell suspension supply syringe 6, an initial filling liquid and extrusion liquid supply syringe 7, and an instruction manual 9. The elements of the administration kit 5 shown in Figs. 1 to 4 and described in the method of use are given the same reference numerals, and detailed description thereof will be omitted.

The instruction manual 9 is an instruction manual to instruct the user to start administration of the cell suspension 3 after filling the administration tube 2 with the entire amount of the cell suspension 3 to be administered in one treatment.

The instruction manual 9 preferably includes at least the following two instructions. The first instruction is to rotate the cell suspension supply syringe 6 back and forth multiple times around the central axis of the cell suspension supply syringe 6 before starting to supply the cell suspension 3 to the administration tube 2 from the cell suspension supply syringe 6 filled with the entire amount of cell suspension 3 to be administered in one treatment. The second instruction is to fill the administration tube 2 with the entire amount of cell suspension 3 by quickly starting to supply the cell suspension 3 to the administration tube 2 from the cell suspension supply syringe 6 after the cell suspension 3 in the cell suspension supply syringe 6 is uniformly dispersed.

The instruction manual 9 may include a third instruction to supply the entire amount of the cell suspension 3 to the administration tube 2 within 2 seconds after the cell suspension 3 is uniformly dispersed. The instruction manual 9 may further include a description that the time for supplying the entire amount of the cell suspension 3 to the administration tube 2 is preferably within 1 second.

As explained in the method of using the administration kit 5 above, if the cell suspension 3 is supplied from the cell suspension supply syringe 6 to the administration tube 2 in a state in which the cell structures 3a in the cell suspension 3 have precipitated in the cell suspension supply syringe 6, the cell structures 3a will clog the tip of the syringe, making it difficult to supply the cell suspension 3 into the administration tube 2. By providing the instruction manual 9, the user can supply the cell structures 3a to the administration tube 2 without clogging the tip of the syringe.

The instruction manual 9 is not limited to being in pamphlet format, and may be included in the administration kit 5A in a form that can be obtained via the Internet from an ID number, barcode, or QR code (registered trademark) assigned to a container or the like of the administration kit 5A.

In the administration kit 5, 5A, the syringe attachment part 28 has only one attachment port for attaching a syringe, but instead of the syringe attachment part 28, a syringe attachment part 34 including a plurality of attachment ports 35 to which a syringe can be attached and a switching valve 36 may be provided (see FIG. 15 described later). The switching valve 36 is disposed between the administration tube 2 and the attachment port 35, and switches the communication state between the administration tube 2 and the plurality of attachment ports 35. That is, the switching valve 36 switches between a state in which the administration tube 2 communicates with the initial filling liquid and extrusion liquid supply syringe 7 and a state in which the administration tube 2 communicates with the cell suspension supply syringe 6. If such a switching valve 36 is provided, it becomes unnecessary to replace the attachment of the syringe. Therefore, the process to be performed before puncturing, such as filling the administration tube 2 with the cell suspension 3, can be performed quickly, and the risk of contamination by bacteria can be reduced.

"Puncture needle"
The puncture needle has a needle body along the front-to-rear direction with a passage for a cell suspension formed therein, and an outlet for the cell suspension formed on the outer surface of the front end of the needle body, the needle body being a single tube with the passage, and the cross-sectional area of the outlet path perpendicular to the direction in which the outlet path extends smoothly increases toward the outlet in at least a part of the outlet path, and at each position in the direction in which the outlet path extends, is greater than or equal to the cross-sectional area at a position closer to the passage.

The puncture needle may have a cross-sectional area of the outlet path that smoothly increases toward the outlet in at least a portion of the outlet path. In other words, at least a portion of the outlet path may expand toward the outlet. Also, the cross-sectional area of the outlet path may be equal to or greater than the cross-sectional area at each position in the direction in which the outlet path extends than at a position closer to the passage. In other words, the cross-sectional area of the outlet path does not change to become smaller toward the outlet. This makes it difficult for the object discharged from the outlet to form a jet. In addition, since the needle body is a single tube with a passage for the object, the outer diameter of the tube can be made smaller than that of a conventional double tube, thereby reducing the burden on the brain and central nervous system. The outer diameter of the needle body may be any. For example, the outer diameter of the needle body is 2.0 mm or less, preferably 1.6 mm or less, and more preferably 1.2 mm or less. Therefore, a puncture needle that is less likely to damage the central nervous system is realized.

In another non-limiting embodiment, it was believed that the jet was formed in the conventional puncture needle because the discharge passage was formed in a straight line, that is, because the cross-sectional area of the discharge passage was constant. Without wishing to be bound by theory, the puncture needle for injecting a fluid object into the central nervous system of the present disclosure has a cross-sectional area of the discharge passage perpendicular to the direction in which the discharge passage extends that smoothly increases toward the discharge outlet in at least a portion of the discharge passage, and at each position in the direction in which the discharge passage extends, is equal to or greater than the cross-sectional area at a position closer to the passage, thereby unexpectedly suppressing or eliminating the jet. This effect is also enhanced by the fact that, in the puncture needle of the present disclosure, which has a needle body along the front-rear direction with a passage for the object formed therein, an object discharge outlet formed on the outer surface of the front end of the needle body, and an object discharge passage extending from the passage to the discharge outlet, the needle body is a single tube with a passage for the object.

It is preferable that the outer surface of the front end portion includes a front end face that protrudes forward while smoothly curving, and a cylindrical side surface connected to the front end face. Conventional puncture needles have a sharp front end or a flat front end face. When such a puncture needle is inserted into a living body, there is a high risk that the needle will damage tissue such as a blood vessel. In contrast, with the above configuration, the front end face is curved, so there is a lower risk that the needle will damage tissue when inserted into the living body.

The puncture needle preferably has the front end of the outlet located behind the front end of the passage, and at the boundary between the front end face and the side face or in front of the boundary. Cells or liquid may remain in the passage in the needle body in front of the outlet and be wasted. In contrast, with the above configuration, the front end of the outlet is located at the boundary between the front end face and the side face or in front of it. Therefore, the portion of the passage from its front end to the outlet passage tends to be relatively short. This results in a relatively small amount of cells or liquid remaining in the portion from the front end of the passage to the outlet passage and being wasted.

It is preferable that the outlet of the puncture needle is formed behind the front end face. In this way, the outlet is hidden behind the front end face when viewed from the front. Therefore, when the puncture needle is inserted forward into the living body, tissue inside the living body is less likely to enter the outlet.

It is preferable that the front end of the puncture needle has a cylindrical side surface, and at least a part of the outlet is formed on the side surface. In this way, since at least a part of the outlet is formed on the side surface, tissue inside the living body is less likely to enter the outlet when the puncture needle is inserted.

The outer surface of the needle body of the puncture needle is the surface that comes into contact with the living body when puncturing, and it is preferable that the wall portion from the passage of the needle body to the outer surface be made of a single layer. This makes it easy to make the inner diameter of the passage as large as possible while keeping the outer diameter of the needle body small. The larger the inner diameter of the passage, the less likely it is that cells etc. will become clogged inside, so it is effective to make the inner diameter of the passage as large as possible.

When water is flowed into the passage from the syringe, it is preferable that the minimum force applied to the puncture needle so that the water discharged from the outlet forms a jet is 1.5 N or more, 2.0 N or more, 2.3 N or more, 2.6 N or more, or 3.0 N or more. This realizes a puncture needle that is less likely to form a jet than conventional needles. In this case, the maximum force applied to the syringe so that the water discharged from the outlet forms a jet can be 6.0 N, 5.7 N, 5.4 N, or 5.0 N.

An example of the puncture needle 1 included in the administration kit 5 (or 5A) will be specifically described with reference to Figs. 81 to 10. As shown in Fig. 8, the puncture needle 1 has a needle body 10 that extends linearly in one direction, and a connection part 20 to which a syringe fixed to the rear end part of the needle body is connected. A cell suspension to be administered into a living body is supplied to the puncture needle 1 from the administration tube 2.

Hereinafter, as shown in Figures 8 to 10, the direction along the needle body 10 is referred to as the front-rear direction. The front is the direction in which the puncture needle 1 is inserted into the living body, and the rear is the opposite direction. Also, as shown in Figures 8, 9, and 10A, a direction perpendicular to the front-rear direction is referred to as the up-down direction. Inside the connection part 20, as shown in Figure 9, there are formed a tube insertion hole 21 into which the first end 2a of the administration tube 2 is inserted, and a body insertion hole 22 into which the needle body 10 is inserted.

The needle body 10 is a circular tubular member made of metal such as stainless steel or aluminum, ceramic, or inflexible hard resin (with little bending). In the present disclosure, the circular shape does not necessarily have to be a strict circle, and may be a shape that is medically acceptable, such as an approximately circular shape, for example, an ellipse. By making the material of the needle body 10 inflexible, such as metal or hard resin, it becomes possible to accurately insert the puncture needle to the desired treatment target point. Preferably, the needle body is made of stainless steel, which is inexpensive and can be easily sterilized. As shown in Figures 9 and 10A, a passage 11 through which cells pass is formed inside the needle body 10. The passage 11 is defined by the inner surface of a through hole that penetrates the needle body 10 in the front-rear direction and the surface of a metal blocking part 12 that blocks the opening at the front end of the through hole. The passage 11 opens toward the rear at the rear end of the needle body 10 and communicates with the tube insertion hole 21 of the connection part 20. The cell suspension supplied from the administration tube 2 is injected into the passage 11 through the tube insertion hole 21. The cells injected into the passage 11 pass through the passage 11 by being pressed from the tube and move toward the front end 13 of the needle body 10. In this embodiment, the needle body 10 is made of a single tube, not a conventional double tube. In other words, the puncture needle 1 is different from the needle described in Patent No. 5665027, which has a double tube structure of an inner needle and an outer needle. Therefore, it is possible to suppress the outer diameter of the needle body 10, thereby reducing the burden on the patient and damage to the central nervous system. In addition, in this embodiment, the wall portion of the needle body 10 from the passage 11 to the outer surface is not divided in the middle and is made of a single layer. In this way, it is possible to form the inner diameter of the passage 11 as large as possible. In addition, the larger the inner diameter of the passage 11, the less likely it is that cells will clog inside. If cells clog the passage 11, the cells and liquid discharged from the discharge port 15 will form a jet as described above, increasing the risk of damaging the brain. Therefore, as described above, it is effective in terms of avoiding the formation of a jet flow to make it easier to make the inner diameter of the passage 11 as large as possible while keeping the outer diameter of the needle body made of a single tube small.

The inner diameter of the passage 11 can be any desired value. For example, the inner diameter is 0.3 mm to 1.2 mm, preferably 0.5 mm to 1.2 mm.

The length of the puncture needle can be any length. For example, the distance between the tip 20a of the connection part 20 and the tip 10a of the needle body 10 is 150 mm to 230 mm, preferably 170 mm to 220 mm, and more preferably 170 mm to 200 mm.

As shown in Figures 10A and 10B, the front end 13 of the needle body 10 has a front end face 13a, which is an outer surface that protrudes forward, and a side face 13b, which is a cylindrical outer surface. The front end face 13a has a smoothly curved hemispherical shape and smoothly connects to the side face 13b at the rear end.

At the front end 13, as shown in FIG. 10B, a circular outlet 15 is formed when viewed from above. Cells supplied from the administration tube 2 into the passage 11 are discharged from the outlet 15. The front end 15a of the outlet 15 is located exactly at the boundary B between the front end face 13a and the side face 13b. In other words, the entire outlet 15 is formed within the range of the side face 13b and is positioned behind the front end face 13a. Therefore, when the puncture needle 1 is viewed from the front, the outlet 15 is hidden behind the front end face 13a and cannot be seen.

Within the front end 13, a cell outlet path 14 is formed, extending upward from the passage 11 to the outlet 15. The outlet path 14 consists of a truncated cone portion 14a, which is approximately truncated cone-shaped, and a cylindrical portion 14b, which is approximately cylindrical. The lower end of the cylindrical portion 14b is connected to the passage 11. The connection position of the cylindrical portion 14b and the passage 11 is a position spaced rearward from the front end of the passage 11. The upper end of the cylindrical portion 14b is connected to the lower end of the truncated cone portion 14a. The lower end of the truncated cone portion 14a has the same shape as the upper end of the cylindrical portion 14b, that is, a circular shape with an inner diameter the same as the inner diameter of the cylindrical portion 14b. The upper end of the truncated cone portion 14a is connected to the cell outlet 15, which opens on the side surface 13b of the front end 13.

The cross section of the outlet path 14 is formed to have the following characteristics. Unless otherwise specified, the cross section of the outlet path 14 is a cross section perpendicular to the direction in which the outlet path 14 extends, that is, a cross section perpendicular to the vertical direction in FIG. 10. The cross section of the outlet path 14 is defined within the range surrounded by the dashed line E in FIG. 10. The dashed line E corresponds to the range excluding the range in which the outlet port 15 is formed and the range in which the opening 14c of the outlet path 14 to the passage 11 is formed. The truncated cone portion 14a is formed so that the cross-sectional area increases smoothly from the lower end to the upper end. The cylindrical portion 14b has a constant cross-sectional area. Therefore, the cross-sectional area of the outlet path 14 consisting of the truncated cone portion 14a and the cylindrical portion 14b at any position in the vertical direction is equal to or greater than the cross-sectional area at a position closer to the passage 11 than that position. For example, the cross section of the truncated cone portion 14a at any position in the vertical direction has a larger area than the cross section at any position on the passage 11 side. Also, the cross section of the cylindrical portion 14b at any position in the vertical direction has the same area as the cross section at any position on the passage 11 side.

According to the present embodiment described above, the outlet path 14 of the puncture needle 1 has the following characteristics. First, in at least a part of the outlet path 14 (specifically, the truncated cone portion 14a), the cross-sectional area increases smoothly toward the outlet 15. In other words, at least a part of the outlet path 14 widens toward the outlet 15. Second, the cross-sectional area of the outlet path 14 at any position in the direction in which the outlet path 14 extends is equal to or greater than the cross-sectional area at a position closer to the passage 11 than that position. In other words, the cross-sectional area of the outlet path 14 does not change so as to become smaller toward the outlet 15. On the other hand, if the cross-section of the outlet path 14 is constant regardless of position, or if the outlet path 14 is formed so that the cross-section becomes smaller (so that the cross-section is narrowed) toward the outlet 15 at any point along the way, the cell suspension discharged from the outlet 15 may form a jet, as shown in the examples described below. In particular, there is a high risk of a jet being formed when it is necessary to increase the pressure with which the cell suspension is injected into the syringe, such as when cells or cell structures become clogged inside the passage 11. If the cell suspension discharged from the outlet 15 forms a jet, there is a high risk of brain damage due to the impact of the jet on the brain. In contrast, the outlet path 14 of the puncture needle 1 has the first and second characteristics as described above. For this reason, as shown in the examples described below, the puncture needle 1 is less likely to form a jet. Therefore, the cell suspension discharged from the outlet 15 is less likely to damage the brain.

In addition, in this embodiment, the front end 13 of the puncture needle 1 has a hemispherical front end surface 13a. On the other hand, if the front end of the needle is sharp or the front end surface is flat, there is a high risk that the needle will damage tissue such as blood vessels when inserted into the brain. In contrast, because the front end surface 13a is curved, there is a low risk that the needle will damage tissue when inserted into the brain.

In addition, in this embodiment, the cell outlet 15 is located within the range of the side surface 13b of the front end 13. Therefore, when the needle body 10 is viewed from the front, the outlet 15 is hidden behind the front end surface 13a and cannot be seen. If the outlet 15 were formed on the front end surface 13a instead of the side surface 13b, the outlet 15 would open toward the front. This increases the possibility that brain tissue will enter the outlet 15 when the puncture needle 1 is inserted into the brain. In contrast, since the outlet 15 is located within the range of the side surface 13b as described above and hidden behind the front end surface 13a, the risk of tissue entering the outlet 15 is reduced.

In addition, in this embodiment, the front end 15a of the outlet 15 is located at the boundary B between the front end surface 13a and the side surface 13b. If the front end 15a of the outlet 15 is located behind this position, the portion from the front end to the outlet path 14 in the passage 11 will be long. Cells that reach this portion may remain in this portion without being discharged from the outlet 15. Therefore, if this portion is long, the amount of cells that are wasted may be large. In contrast, as described above, since the front end 15a of the outlet 15 is located at the boundary B between the front end surface 13a and the side surface 13b, the length of the portion from the front end of the passage 11 to the outlet path 14 is relatively small. Therefore, the length of the puncture needle inserted into the central nerve can be shortened as much as possible, so that the burden on the patient and the damage to the central nerve can be reduced. In addition, the amount of cells that are wasted is likely to be relatively small. From the viewpoint of minimizing the length of the portion from the front end of the passage 11 to the outlet passage 14, it is preferable to position the rear end of the blocking portion 12 as far back as possible, that is, as close to the outlet passage 14 as possible. Also, from the viewpoint above, it is most preferable to position the rear end of the blocking portion 12 at the same position as the front end of the cylindrical portion 14b.

In the embodiment shown in FIG. 10, the angle between one inclined surface and the other inclined surface of the truncated cone portion 14a (cone angle) is approximately 90°, but this cone angle is not limited to 90°. A range of 45° to 150° is preferable, 70° to 110° is more preferable, 85° to 95° is even more preferable, and 88° to 92° is even more preferable. The inner diameter of the cylindrical portion 14b can be any size as long as it is equal to or smaller than the inner diameter of the cylindrical portion 14b. For example, the inner diameter of the cylindrical portion 14b is 0.3 mm to 1.2 mm. The inner diameter of the outlet 15 can be any size as long as it is equal to or smaller than the diameter of the needle body 10. For example, the inner diameter of the outlet 15 is 1.2 mm to 1.6 mm.

In this puncture needle 1, the outlet path 14 is provided along the vertical direction, but may be provided so as to extend from the passage 11 to the outlet 15 along a direction inclined forward at an angle θ with respect to the vertical direction. The inclination angle θ with respect to the vertical direction can be any angle. For example, it is greater than 0° and 30°, and preferably 10° to 20°. In this case, it is preferable that the minimum inner diameter of the outlet path is 0.3 mm to 1.2 mm, and the inner diameter of the outlet is 0.3 mm to 1.2 mm.

<Example of needle design change>
An example of a modified design of the puncture needle is shown in Figure 11. The administration kit 5 of the present disclosure may include a puncture needle 1C shown in Figure 11 instead of the puncture needle 1. In Figure 11, elements equivalent to the puncture needle 1 shown in Figures 8 to 10 are denoted by the same reference numerals, and detailed description thereof will be omitted.

The puncture needle 1C has a needle body 10 with a passage 11 for the cell suspension 3 formed therein, and a connection part 20 that holds the base end side of the needle body 10. The first end 2a of the administration tube 2 is attached to the connection part 20. The connection part 20 has a body insertion hole 22 into which the base end side of the needle body 10 is inserted, and a tube insertion hole 21 into which the first end 2a of the administration tube 2 is inserted. The body insertion hole 22 and the tube insertion hole 21 are connected, and the cell suspension 3 discharged from the administration tube 2 is supplied to the passage 11 of the needle body 10. The puncture needle 1C has an outlet 15 for the cell suspension 3 on the outer surface of the front end 13 of the needle body 10.

As shown in FIG. 11, the tip end portion 21a that connects to the main body insertion hole 22 of the tube insertion hole 21 of the connection part 20 has a funnel shape with an inner diameter that gradually narrows from the base end toward the tip end. In other words, the tip end portion 21a has a shape that is tapered from the base end toward the tip end. The opening angle α of the funnel shape that opens from the tip end toward the base end (the opening angle of the tapered surface 25 in the cross-sectional view of FIG. 11) is preferably 120° or less.

In this puncture needle 1C, the tip portion 21a of the tube insertion hole 21 is funnel-shaped, which allows the cell structure 3a to be smoothly introduced into the passage of the needle body 10. It is preferable that the difference in inner diameter between the body insertion hole 22 and the tube insertion hole 21 at the boundary between them is small, and it is particularly preferable that the inner diameters of both are the same. The smaller the difference in inner diameter between the body insertion hole 22 and the tube insertion hole 21 at the boundary between them, the smaller the step where the cell structure 3a will be retained will be, and this can prevent the cell structure 3a from getting caught on the step and becoming clogged.

The puncture needle 1C preferably has a diameter of 0.5 mm or more in the passage 11 of the needle body 10. A diameter of 0.5 mm or more can more effectively prevent clogging of cell structures 3a of a certain size, for example, a diameter of about 300 μm or less.

The diameter of the passage of the needle body 10 of the puncture needle 1C (the inner diameter φ1 of the needle body 10) is preferably at least twice the diameter of the cells or cell structures 3a contained in the cell suspension, more preferably at least three times, and even more preferably at least four times. If the diameter of the passage 11 of the needle body 10 is at least twice the diameter of the cells or cell structures 3a, the cell structures 3a can be administered to a living body without clogging the cells or cell structures 3a inside the passage. The inner diameter φ1 of the needle body 10 of the puncture needle 1C is preferably at least 0.5 mm, more preferably at least 0.8 mm, and even more preferably at least 1.0 mm.

The puncture needle 1C has an outer diameter of the needle body 10 of preferably 3 mm or less, more preferably 2 mm or less, and even more preferably 1.5 mm or less. By making the outer diameter of the needle body 10 3 mm or less, damage to the living body being punctured can be suppressed.

The puncture needle 1C preferably has an opening diameter φ3 of the outlet 15 that is equal to or greater than the diameter φ1 of the passage 11 of the needle body 10. As shown in the passage 11 of the needle body 10, if the opening diameter φ3 of the outlet 15 is equal to or greater than the diameter φ1 of the passage 11 of the needle body 10, the cell structure 3a can be passed through smoothly.

It is preferable that the opening area of the outlet 15 of the puncture needle 1C is equal to or larger than the cross-sectional area of the passage 11 of the needle body 10. If the opening area of the outlet 15 is equal to or larger than the cross-sectional area of the passage 11 of the needle body 10, the cell structure 3a can be passed through smoothly.

The puncture needle 1C preferably has a needle body 10 length of 100 mm or more. For example, it is suitable for transplanting cells such as stem cells to a specific location in the brain in stereotactic brain surgery. A cell suspension can be administered to the affected area even if the affected area is located deep below the surface of the head. For example, a tube length of 190 mm is a suitable length when performing cell transplant surgery using a stereotactic brain surgery device.

Below, examples and comparative examples of the administration kit of the present disclosure will be described. First, the method for producing the cell structure will be described.

<Method of producing cell structure>
((Recombinant peptide (recombinant gelatin))
The following CBE3 was prepared as a recombinant peptide (recombinant gelatin) (described in WO 2008/103041).
CBE3:
Molecular weight: 51.6kDa
Structure: GAP[(GXY)63]3G
Number of amino acids: 571 RGD sequences: 12 Imino acid content: 33%
Almost 100% of the amino acids are repeat structures of GXY. The amino acid sequence of CBE3 does not contain serine, threonine, asparagine, tyrosine, or cysteine. CBE3 has the ERGD sequence.
Isoelectric point: 9.34
GRAVY value: -0.682
1/IOB value: 0.323
Amino acid sequence (SEQ ID NO: 1 in the sequence listing) (same as SEQ ID NO: 3 in WO 2008/103041, except that the final X has been changed to "P")
GAP GERGAAGLPGPKGERGDAGPKGADGAPGKDGVRGLAGPIGPPGPAGAPGAPGLQGMPGERGAAGLPGPKGERGDAGPKGADGAPGKDGVRGLAGPP) _3G

(Preparation of recombinant peptide porous material)
[Thick PTFE cylindrical container]
A cylindrical cup-shaped container made of polytetrafluoroethylene (PTFE) was prepared, with a bottom thickness of 3 mm, a diameter of 51 mm, a side thickness of 8 mm, and a height of 25 mm. When the cylindrical cup has a curved surface as a side, the side is closed with 8 mm of PTFE, and the bottom (flat circular plate) is also closed with 3 mm of PTFE. On the other hand, the top surface is open. Therefore, the inner diameter of the cylindrical cup is 43 mm.

[Freezing process and drying process with small temperature difference]
The CBE3 aqueous solution was poured into a PTFE thick cylindrical container, and cooled from the bottom using a cooling shelf in a vacuum freeze dryer (TF5-85ATNNN: Takara Seisakusho). At this time, the final concentration of the CBE3 aqueous solution was 4 mass%, and the amount of the aqueous solution was 4 mL. The shelf temperature was set to -10°C, and the solution was frozen at -10°C for 1 hour, then at -20°C for 2 hours, then at -40°C for 3 hours, and finally at -50°C for 1 hour. The frozen product was then vacuum-dried at -20°C for 24 hours after the shelf temperature was returned to -20°C, and after 24 hours, the shelf temperature was raised to 20°C while continuing the vacuum drying, and vacuum-dried at 20°C for 48 hours until the degree of vacuum was sufficiently reduced (1.9 x 10 ⁵ Pa), and then removed from the vacuum freeze dryer. A CBE3 porous body was obtained as described above.

(Preparation of biocompatible polymer blocks (pulverization and crosslinking of porous bodies))
The CBE3 porous body obtained by the above procedure was pulverized in a New Power Mill (Osaka Chemical, New Power Mill PM-2005). The pulverization was performed at the maximum rotation speed for 1 minute x 5 times, for a total of 5 minutes. The pulverized material obtained was sized using a stainless steel sieve to obtain uncrosslinked blocks of 53 to 106 μm. This was then subjected to thermal crosslinking (48 hours) at 160° C. under reduced pressure to obtain a biocompatible polymer block (CBE3 block).

(Preparation of cell structures)
Human bone marrow-derived mesenchymal stem cells (hMSCs) were suspended in a growth medium (Takara Bio: MSCGM BulletKit™), and the biocompatible polymer block obtained by the above procedure was added thereto. As a result, 6.9 x ¹⁰⁶ cells of hMSCs and 5.76 mg of the biocompatible polymer block were finally suspended in 23 mL of growth medium, and seeded in a 90 mm diameter dish-type culture vessel having multiple depressions on the bottom surface.
After leaving the cells in a _CO2 incubator at 37°C for 3 days, multiple cell structures were collected as spheres with a diameter of about 250 μm consisting of hMSCs and biocompatible polymer blocks. The number of cell structures was measured using a high-speed 3D cell scanner (Cell3iMager neo: SCREEN Holdings). Eight 90 mm dishes were collected, and approximately 55,000 cell structures were obtained.

In the following examples and comparative examples, it was assumed that 3 mL or less of cell suspension 3 containing approximately 55,000 cell structures 3a with an average particle size of 250 μm would be injected into the affected area of the brain. The goal was to administer the cell structure 3a at a 45-degree needle position using a puncture needle 1 with a tube length (length of needle body 10) of 190 mm and a tube outer diameter (outer diameter φ2 of needle body 10) of 1.5 mm, with a puncture time of 10 minutes and administration time of 3 minutes. The average particle size of cell structures 3a was determined by measuring the particle diameter of cell structures 3a using the high-speed 3D cell scanner described above.

[Comparative Example]
Approximately 55,000 of the cell structures obtained by the above procedure were mixed with ArtCeleb (a cerebral and spinal surgery irrigation and perfusion solution (Otsuka Pharmaceuticals)) in a 5 mL syringe (Terumo 5 mL SS-05SZ) to prepare 3 mL of cell suspension. This resulted in a cell suspension supply syringe filled with 3 mL of cell suspension.

The comparative example is an example of a case where an administration kit without an administration tube 2 is used. The administration kit of the comparative example includes a puncture needle 1 and a cell suspension supply syringe 6 containing 3 mL of cell suspension 3.

The puncture needle 1A shown in Figure 12 was used as the puncture needle 1. The needle body 10 of the puncture needle 1A has an inner diameter φ1 of 0.5 mm, an outer diameter φ2 of 1.5 mm, and a tube length of 190 mm. The outlet 15 for the cell suspension of the needle body 10 is located on the side of the front end portion 13, and its hole diameter φ3 is 0.5 mm, the same as the inner diameter φ1 of the needle body 10. The unit of dimensions in Figure 12 is mm. Note that in Figure 12, elements equivalent to the puncture needle 1 shown in Figures 8 to 11 are given the same reference numerals.

To prevent damage to the brain when puncturing it, it is desirable for the outer diameter φ2 of the needle body 10 to be as small as possible, so the outer diameter was set to 1.5 mm. Also, to enable administration to the affected area even if it is deep below the surface of the head, the tube length was set to 190 mm.

The procedure for administering the cell suspension using this administration kit was as follows:

A cell suspension supply syringe 6 filled with cell suspension 3 was inserted into the puncture needle 1A. With the cell suspension supply syringe 6 and the puncture needle 1A in a horizontal position, the cell suspension supply syringe 6 and the puncture needle 1A were repeatedly rotated back and forth around the central axis of the cell suspension supply syringe 6. As a result, the cell suspension 3 in the cell suspension supply syringe 6 became in a state in which the cell structures 3a were uniformly dispersed.

The puncture needle 1A was quickly moved to a position at 45° from horizontal, which is the position assumed for administration to the brain, and the cell suspension 3 was immediately pushed out from the cell suspension supply syringe 6. When 3 mL of cell suspension was pushed out in a short time of about 1 second, the cell suspension 3 could be pushed out from the tip of the puncture needle.

In actual treatment, it takes about 10 minutes to puncture the brain. In addition, the drug must be injected slowly into the brain over a period of time; for example, 3 mL must be injected slowly over a period of 3 minutes.

Figure 13 shows photographs taken every 5 seconds of the cell structures sinking with the needle at a 45° position, with the cell suspension in the syringe uniformly dispersed. 0 seconds is the state immediately after the cell suspension is uniformly dispersed. As shown in Figure 13, after 5 seconds, it can be seen that part of the cell suspension, about the top 1/4 of the cell suspension, has become transparent, and after 10 seconds, about 1/2 of the cell suspension in the syringe has become transparent. After 20 seconds, the cell structures in the cell suspension have completely sunk.

In this way, if a cell suspension in which the cell structures are uniformly dispersed in the syringe is left for the duration of the puncture and long administration time, the cell structures in the syringe will settle and will no longer be able to remain suspended. When an attempt is made to push out the cell structures in the cell suspension supply syringe in this state in which the cell structures have settled in the cell suspension, the cell structures become clogged at the tip of the syringe or at the entrance to the puncture needle. It is believed that because the settled cell structures have no fluidity, their movement is inhibited, resulting in the clogging.

[Example 1]
Approximately 55,000 cell structures obtained by the above procedure were mixed with ArtCeleb (a cerebral and spinal cord surgery irrigation and perfusion fluid, manufactured by Otsuka Pharmaceuticals) in a 5 mL syringe (Terumo 5 mL SS-05SZ) to prepare 2.4 mL of cell suspension. This resulted in a cell suspension supply syringe 6 filled with 2.4 mL of cell suspension 4. Note that ArtCeleb is a liquid that is also used as the initial filling liquid and extrusion liquid in the following examples, and is an example of artificial cerebrospinal fluid 8. In the following, artificial cerebrospinal fluid 8 refers to ArtCeleb.

The administration kit of this example includes a puncture needle 1, an administration tube 2, and a cell suspension supply syringe 6 filled with the above-mentioned cell suspension 3. This example will be described with reference to Figures 2 to 5. Elements equivalent to those described with reference to Figures 2 to 5 will be described with the same reference numerals. Furthermore, in this example, the puncture needle 1A shown in Figure 12 was used as the puncture needle 1. The puncture needle 1 in Figures 2 to 5 will be described as puncture needle 1A.

An initial filling liquid and extrusion liquid supply syringe (Terumo 10 mL SS-10LZ) 7 used as an initial filling liquid supply and extrusion liquid supply syringe was filled with 10 mL of artificial cerebrospinal fluid 8.

As the puncture needle, the puncture needle 1A shown in FIG. 12 was used as in the comparative example.
As the administration tube 2, a spiral tube having an inner diameter of 1.1 mm and a length of 3 m (manufactured by TOP, X1-300S, capacity 2.5 mL) was used.

The procedure for administering the cell suspension using this administration kit was as follows:

First, the puncture needle 1A, the administration tube 2, and the initial filling liquid and extrusion liquid supply syringe 7 were connected (see FIG. 2).
The puncture needle 1A was held in a position tilted 45 degrees from the horizontal. By supplying about 5 mL of artificial cerebrospinal fluid 8 from the initial filling liquid and extrusion liquid supply syringe 7, the puncture needle 1A and the administration tube 2 were filled with the artificial cerebrospinal fluid 8 and were free of air bubbles. The initial filling liquid and extrusion liquid supply syringe 7 was removed from the administration tube 2, and the cell suspension supply syringe 6 was connected (see FIG. 3).

Next, the cell suspension supply syringe 6 was repeatedly rotated back and forth around the central axis of the syringe to uniformly disperse the cell structures 3a in the cell suspension in the cell suspension supply syringe 6, and the entire amount was supplied to the administration tube 2 in a short period of time (approximately 1 second). This operation resulted in the cell suspension in the cell suspension supply syringe 6 being in a state in which the cell structures 3a were uniformly dispersed along the length of the tube (see Figure 4).

Next, the cell suspension supply syringe 6 was removed from the administration tube 2, and the initial filling liquid and extrusion liquid supply syringe 7 was reconnected (see FIG. 5). After that, it was left for 10 minutes, which is the time required for puncturing the brain. During this 10-minute period, the cell structures 3a in the administration tube 2 hardly moved and maintained a state of being uniformly dispersed in the longitudinal direction of the administration tube 2.

Next, 0.2 mL was extruded from the initial filling liquid and extrusion liquid supply syringe 7 in about 1 second. The flow rate at this time was about 12 mL/min (0.2 x 60/1 = 12). At this time, the cell structure 3a in the administration tube 2 moved inside the administration tube 2 while maintaining a uniformly dispersed state in the length direction, and was left for 11 seconds, and 0.2 mL was again extruded from the initial filling liquid and extrusion liquid supply syringe 7 in about 1 second. This extrusion and leaving operation was repeated multiple times.

When the cell suspension 3 was extruded from the tip of the puncture needle using the above administration procedure, the cell structures 3a could be extruded from the tip of the puncture needle without any problems, at least up to the fourth extrusion after the start of extrusion. In the comparative example, when the administration tube 2 was not provided, the cell structures 3a precipitated at the tip of the cell suspension supply syringe 6 left for 10 minutes, and the cell structures 3a became clogged at the tip of the cell suspension supply syringe 6. However, by using the administration tube 2 as in this example, clogging could be suppressed. This is thought to be because the cell structures 3a in the cell suspension in the administration tube were distributed almost uniformly inside. However, in this example, the cell structures 3a became clogged at the entrance of the puncture needle 1A after the fifth extrusion.

[Example 2]
As shown in FIG. 14, the administration kit of Example 2 includes a puncture needle 1, an administration tube 2, and a cell suspension supply syringe 6 filled with a cell suspension 3. In this example, the puncture needle 1A shown in FIG. 12 was used as the puncture needle 1. The administration tube 2 and the cell suspension supply syringe 6 filled with a cell suspension 3 were the same as those in Example 1. Furthermore, one initial filling liquid supply syringe (10 mL SS-10LZ manufactured by Terumo Corporation) 7A and three extrusion liquid supply syringes (1 mL SS-01T manufactured by Terumo Corporation) 7B were provided. That is, the administration kit of Example 2 differs from Example 1 in that an initial filling liquid supply syringe and an extrusion liquid supply syringe are provided, whereas Example 1 includes an initial filling liquid supply and an extrusion liquid supply syringe that serve both initial filling supply and extrusion liquid supply.

As shown in FIG. 14, the inner diameter of the extrusion liquid supply syringe 7B is smaller than that of the initial filling liquid supply syringe 7A. By using the extrusion liquid supply syringe 7B with a smaller inner diameter, the cell structure cell suspension can be extruded at a higher pressure. The pressure generated in the syringe and administration tube 2 is inversely proportional to the cross-sectional area. Since the cross-sectional area of a 1 mL syringe is smaller than that of a 10 mL syringe, it is possible to extrude at a higher pressure. In addition, since the stroke when extruding 0.2 mL at a time is longer than that of a 10 mL syringe, it is possible to extrude quickly in a short period of time.

The procedure for administering the cell suspension using this administration kit was as follows:

First, as shown in FIG. 14, the puncture needle 1A, the administration tube 2, and the initial filling liquid supply syringe 7A were connected.

The puncture needle 1A was held in a position tilted at approximately 45 degrees from the horizontal. By supplying 5 mL of artificial cerebrospinal fluid 8 from the initial filling fluid supply syringe 7A, the puncture needle 1A and administration tube 2 were filled with artificial cerebrospinal fluid 8, and no air bubbles were present.

Next, the initial filling liquid supply syringe 7A was removed from the administration tube 2, and the cell suspension supply syringe 6 was connected. At this time, care was taken to avoid introducing air bubbles into the connection.

Next, the cell suspension supply syringe 6 was repeatedly rotated back and forth around the central axis of the syringe to uniformly disperse the cell structures 3a in the cell suspension 3 in the cell suspension supply syringe 6, and the entire amount was supplied to the administration tube 2 in a short time (about 1 second). This operation resulted in the cell suspension 3 in the cell suspension supply syringe 6 being uniformly dispersed in the longitudinal direction of the administration tube 2.

Next, the cell suspension supply syringe 6 was removed from the administration tube 2, and the extrusion liquid supply syringe 7B was connected. After that, it was left for 10 minutes, which is the time required for puncturing the brain. During this 10-minute period, the cell structures 3a in the administration tube 2 hardly moved, and maintained a state of being uniformly dispersed in the longitudinal direction of the administration tube 2.

Next, 0.2 mL was extruded from the extrusion liquid supply syringe 7B in about 0.5 seconds. The flow rate at this time was about 24 mL/min (0.2 x 60/0.5 = 24). This extrusion caused the cell structure 3a in the administration tube 2 to move within the tube 2 while remaining uniformly dispersed in the longitudinal direction. After this, the solution was left to stand for about 11.5 seconds, and again 0.2 mL was extruded from the extrusion liquid supply syringe 7B in about 0.5 seconds. This extrusion and leaving operation was repeated five times.

The first extrusion liquid supply syringe 7B, which had become empty as a result of the above operation, was removed from the administration tube 2, and the second extrusion liquid supply syringe 7B was attached. As with the first extrusion liquid supply syringe 7B, the extrusion and leaving operation was repeated five times. Then, the second extrusion liquid supply syringe 7B, which had become empty, was removed from the administration tube 2, and the extrusion and leaving operation was repeated five times with the third extrusion liquid supply syringe 7B, as described above.

A total of 15 extrusions were performed over the course of 3 minutes using the above series of operations, resulting in a total of 3 mL extrusion. As a result, almost the entire amount was able to be extruded from the tip of the puncture needle without clogging the cell structure 3a. In other words, it became clear that the entire amount of cell suspension to be administered in one treatment could be administered without clogging.

In this example, the puncture needle 1A and administration tube 2 are filled with artificial cerebrospinal fluid 8 in advance, and the entire amount of cell suspension 3 in the cell suspension supply syringe 6 is supplied to the administration tube 2 in a short period of time while the cell suspension 3 is uniformly dispersed. After that, the cell structure 3a is extruded at high pressure and high speed using the extrusion liquid supply syringe 7B, which has a small inner diameter, thereby preventing clogging of the cell structure 3a.

[Example 3]
As shown in Fig. 15, the administration kit of Example 3 includes a puncture needle 1, an administration tube 2, a cell suspension supply syringe 6 filled with a cell suspension 3, an initial filling liquid and extrusion liquid supply syringe 7, and a connection tube 31. In this example, the puncture needle 1A shown in Fig. 12 was used as the puncture needle 1. In this example, the syringe attachment part 34 at the second end 2b of the administration tube 2 includes two syringe attachment ports 35 and a switching valve (3W-RC manufactured by Nipro) 36.

A cell suspension supply syringe 6 was attached to one of the syringe mounting ports 35, and an initial filling liquid and extrusion liquid supply syringe 7 was attached to the other via a connection tube 31.

The switching valve 36 can be switched between a state in which the administration tube 2 communicates with the cell suspension supply syringe 6 and a state in which the administration tube 2 communicates with the initial filling liquid and extrusion liquid supply syringe 7.

The connection tube 31 is a tube that connects the attachment port 35 to the initial filling liquid and extrusion liquid supply syringe, and is a Nipro EX1-50NFH with a capacity of 0.5 mL.

According to the configuration of this example, in the first embodiment, it is possible to switch between the supply of artificial cerebrospinal fluid and the supply of cell suspension by switching using the switching valve 36 without replacing the syringe.
Compared to the configurations of Examples 1 and 2, there is no need to replace the syringe, which improves operability and reduces the risk of air bubbles or foreign matter being mixed in.

[Example 4]
As shown in FIG. 16, the administration kit of Example 4 is the administration kit of Example 3, except that instead of the initial filling liquid and extrusion liquid supply syringe 7, it includes one initial filling liquid supply syringe (10 mL SS-10LZ, manufactured by Terumo Corporation) 7A and three extrusion liquid supply syringes (1 mL SS-01T, manufactured by Terumo Corporation) 7B.

When supplying the extrusion liquid, the extrusion liquid supply syringe 7B was replaced in the same manner as in Example 2. Otherwise, the administration procedure was the same as in Example 2, and as in Example 2, no clogging occurred and almost the entire amount was able to be dispensed from the tip of the puncture needle 1A.

[Example 5]
As shown in Figure 17, the administration kit of Example 5 is the administration kit of Example 4, in which an attachment portion 44 for attaching a syringe to a connecting tube 31 has attachment ports 45 for an initial filling liquid supply syringe 7A and three extrusion liquid supply syringes 7B, and has three second switching valves (16901C manufactured by B. Braun) 46 that switch the flow path to connect any of the syringes to the connecting tube 31.

By providing the second switching valve 46, the work of replacing the extrusion liquid supply syringe 7B is eliminated. When replacing the extrusion liquid supply syringe 7B as in Example 4, careful operation is required to prevent the introduction of foreign matter such as air bubbles and germs. However, since there is no replacement work, high operability can be obtained without the risk of introducing air bubbles or foreign matter.

The initial filling liquid supply syringe 7A and the extrusion liquid supply syringe 7B were connected to the second switching valve 46 in advance, and when using each syringe, the second switching valve 46 was switched and used. As with Example 2, no clogging occurred when this administration kit was used, and almost the entire amount could be dispensed from the tip of the puncture needle 1A.

In a configuration such as Example 5 where the syringe replacement work is not required, stable administration is possible without the risk of air bubbles or foreign matter being mixed in. On the other hand, the configuration becomes complicated as multiple switching valves are required, and a separate check of the switching operation is required. Therefore, the administration conditions under which the cell suspension can be reliably administered in a simple configuration such as Example 3 (see Figure 15) were examined as follows.

(Consideration of administration conditions)
Specifically, in the configuration of Example 3, by changing the inner diameter of the needle body 10 of the puncture needle 1, the connection structure, and the inner diameter and length of the administration tube 2, we investigated conditions under which administration can be performed without requiring high pressure.

Evaluation 1) Inner diameter of needle body 10 of puncture needle 1 Puncture needle 1A shown in Fig. 12 and puncture needle 1B shown in Fig. 19 were prepared. For puncture needle 1B in Fig. 19, the same elements as those of needle 1A are given the same reference numerals. The inner diameter φ1 of the needle body 10 of puncture needle 1A is 0.5 mm, and the hole diameter φ3 of the outlet is 0.5 mm, whereas the inner diameter φ1 of the needle body 10 of puncture needle 1B is 1.10 mm, and the hole diameter φ3 of the outlet is 1.1 mm. Other than the inner diameter of the needle body 10 and the hole diameter of the outlet, puncture needle 1A and puncture needle 1B are the same.

Using the method described below, the maximum cell structure concentration (pieces/mL) at which cell structures 3a having an average particle size of 250 μm can be extruded using the configuration of Example 3 was obtained.
The initial filling liquid and extrusion liquid supply syringe 7 was filled with 10 mL of artificial cerebrospinal fluid 8 .
Approximately X cell structures were mixed with 2.4 mL of artificial cerebrospinal fluid 8 to prepare a cell suspension. At this time, X was changed to prepare a number of cell suspensions with different concentrations Y of cell structures 3a in the cell suspension, as shown in Table 1. When the number of cell structures is X, the cell structure concentration Y in the cell suspension is:
Y=X/2.4 (pcs/mL)
It is.

An evaluation was performed to see whether the cell suspension 3 could be completely dispensed from the tip of the puncture needle 1 using the following administration procedure. The procedure was performed in order starting from the lowest cell structure concentration in Table 1. The maximum concentration at which the entire amount of cell structures could be dispensed from the end of the puncture needle 1A without clogging was determined as the limit concentration.

The administration procedure was as follows:

The puncture needle 1A was held at a 45-degree angle. The switching valve 36 was switched to a state in which the administration tube 2 was in communication with the initial filling liquid and extrusion liquid supply syringe 7, and approximately 5 mL of artificial cerebrospinal fluid 8 was supplied from the initial filling liquid and extrusion liquid supply syringe 7, filling the puncture needle 1A and administration tube 2 with the artificial cerebrospinal fluid 8 and eliminating any air bubbles.

Next, the switching valve 36 was switched to a state in which the administration tube 2 and the cell suspension supply syringe 6 were in communication.

Next, the cell suspension supply syringe 6 was repeatedly rotated back and forth around the central axis of the syringe to uniformly disperse the cell structures 3a of the cell suspension 3 in the cell suspension supply syringe 6, and the entire amount was supplied to the administration tube 2 in a short time (about 1 second). This operation resulted in the cell suspension in the cell suspension supply syringe 6 being uniformly dispersed in the longitudinal direction of the administration tube 2.

Next, the switching valve 36 was switched so that the administration tube 2 and the cell suspension supply syringe 6 were connected, and the tube was left for 10 minutes, which corresponds to the time required for puncturing the brain. After that, 0.2 mL of artificial cerebrospinal fluid 8 was extruded from the initial filling liquid and extrusion liquid supply syringe 7 in about 1 second. At this time, the cell structure 3a in the administration tube 2 moved within the administration tube 2 while maintaining a uniformly dispersed state in the length direction. After that, the tube was left for 11 seconds, and 0.2 mL of artificial cerebrospinal fluid 8 was again extruded from the initial filling liquid and extrusion liquid supply syringe 7 in about 1 second. This extrusion and leaving operation was repeated a total of 15 times. The time required for extrusion was about 3 minutes.

The results are shown in Table 2.

The results showed that increasing the inner diameter of the needle body 10 increased the limiting concentration. In other words, it was found that a larger inner diameter of the needle made it less likely to become clogged. This is thought to be because by increasing the inner diameter of the needle, it became less likely for the cell structure 3a to become blocked at the entrance of the needle body 10 or within the passage of the needle body 10.

Evaluation 2) Shape of the connecting part of the puncture needle 1 The shape of the tube insertion hole 21 of the connecting part 20 of the puncture needle 1 was changed to confirm the effect.
The puncture needle 1B shown in FIG. 18 and the puncture needle 1C shown in FIG. 19 were prepared. In both cases, the inner diameter φ1 of the needle body 10 was 1.1 mm. In the puncture needle 1C shown in FIG. 19, the tip end portion of the tube insertion hole 21 of the connection part 20 is funnel-shaped. That is, a tapered surface 25 is provided at the tip end portion of the tube insertion hole 21. The opening angle α of the funnel shape is 120°. The difference between the inner diameter of the needle body 10 and the tip end portion 21a closest to the needle body 10 is 0.1 mm. In contrast, the puncture needle 1B shown in FIG. 18 does not have a funnel-shaped tip end portion 21a at the connection part 20.

The limit concentration for the puncture needles 1B and 1C was evaluated using the same evaluation method as above. The results are shown in Table 3. It was found that tapering the tip portion 21a of the insertion hole 21 increased the limit concentration, i.e., made it less likely to become clogged. This is thought to be because tapering the tip portion 21a of the insertion hole 21, which serves as the entrance to the needle body 10, allowed the cell structure 3a to flow smoothly into the needle body due to the water flow of the artificial cerebrospinal fluid 8.

Evaluation 3) Inner Diameter of Administration Tube When the above-mentioned puncture needle 1C was used, the effect was confirmed for administration tubes 2 having different inner diameters.
As administration tubes 2, ones with an inner diameter of 1.1 mm, a length of 3 m, and a capacity of 2.4 mL (manufactured by Top Co., Ltd.) and ones with an inner diameter of 1.5 mm, a length of 1.5 m, and a capacity of 2.7 mL (manufactured by Bigon Co., Ltd., 1159.65) were prepared.
The limit concentration was evaluated using two types of administration tubes 2 using the same evaluation method as in Evaluation 1).
The results are shown in Table 4.

It was found that the limit concentration was higher, that is, the administration tube 2 having an inner diameter of 1.5 mm and a length of 1.5 m was less likely to become clogged, than the administration tube 2 having an inner diameter of 1.1 mm and a length of 3 m.
It is believed that by increasing the inner diameter and shortening the length of the administration tube 2, the pressure loss in the administration tube 2 during extrusion is reduced, and as a result, the pressure near the entrance from the administration tube 2 to the needle body 10 becomes higher, making it less likely to become clogged.

The results of evaluations 1 to 3 are summarized in Table 5.
The limit concentration ratios in Table 5 are concentration ratios relative to the target concentration (the concentration when 55,000 cell structures are suspended in 2.4 mL of artificial cerebrospinal fluid 8, i.e., 55,000/2.4=22,917 cells/mL) that was the target concentration of the study.

By changing the inner diameter (needle inner diameter) of the needle body 10, the shape of the insertion hole that connects to the base end of the needle body 10, and the inner diameter and length of the administration tube, it was possible to stably dispense a highly concentrated cell suspension, approximately 2.3 times the concentration of the target, from the tip of the puncture needle without clogging.

Here, under the above four conditions, the puncture needle 1, the administration tube 2, and the initial filling liquid and extrusion liquid supply syringe 7 were filled with water at 25°C, and the syringe pressing force required to push out 0.2 mL in 1 second was measured. The syringe pressing force was measured by pushing the syringe through a push-pull gauge.

The results are shown in Table 6.

At level d, where a high-concentration cell suspension could be stably extruded from the tip of the puncture needle without clogging, the pushing force required to extrude 0.2 mL in 1 second was as small as 4.0 (N), and the operability of extrusion was also good.

[Example 6]
Using an administration kit having a configuration of level d in the above administration condition study, the conditions at the time of the above evaluation were set as standard conditions, and more severe conditions were set with respect to the standard conditions, and an evaluation was made as to whether the cell structure 3a could be removed from the tip of the puncture needle without clogging. The conditions shown in Table 7 were set as the severe conditions. A needle attitude of 90° means a state in which the needle is maintained in the vertical direction. In the extrusion pattern, 15 times with a 12-second interval means that the cell suspension is extruded for 1 second and left for 11 seconds, for a total of 12 seconds, is repeated 15 times. The "Explanation" item in Table 7 explains the severe conditions.

The following conditions were common to all the tests.
Number of cell structures: 55,000 Extrusion: 0.2 mL extruded per second

As a result of the above harsh conditions, when the evaluation of harsh conditions x, y, and z was performed individually, and when the three harsh conditions x, y, and z were performed simultaneously, the cell structure 3a was able to be ejected from the tip of the puncture needle without clogging.

[Example 7]
In the administration kit having the configuration of level d in the above administration condition study, an evaluation was performed to see whether administration was possible under standard conditions using a puncture needle with an inner diameter of 0.8 mm in the needle body 10. The configuration of the administration kit in this example is the same as that in Example 3 (FIG. 15), but the conditions of the puncture needle and administration tube are different from those in Example 3.
As a result of evaluation under standard conditions, it was found that almost all of the cell structures 3a could be ejected from the tip of the puncture needle without clogging.

As described above, since it is difficult to maintain a uniform suspension state in the cell suspension supply syringe for a long period of time, there is a problem that it is difficult to administer the cell structures 3a without clogging them. In contrast, as in Examples 1 to 7, by providing a dosing tube 2 capable of holding the entire amount of cell suspension 3 to be administered in one treatment between the puncture needle and the cell suspension supply syringe, administration was possible without clogging the cell structures 3a. With the puncture needle 1, dosing tube 2, and cell suspension supply syringe 6 connected, the cell suspension supply syringe 6 is repeatedly rotated back and forth around the central axis of the syringe to make the cell suspension 3 in the syringe 6 uniformly dispersed, and the entire amount is supplied to the dosing tube 2 in a short time (about 1 second). After that, by intermittently supplying the initial filling liquid and the extrusion liquid from the extrusion liquid supply syringe 7, it was possible to extrude almost the entire amount of the cell structures 3a from the tip of the puncture needle.

Reference Signs List 1, 1A, 1B, 1C Puncture needle 2 Administration tube 2 Tube 2a First end 2b Second end 3 Cell suspension 3a Cell structure 4 Container 5, 5A Administration kit 6 Cell suspension supply syringe 7 Initial filling liquid and extrusion liquid supply syringe 7A Initial filling liquid supply syringe 7B Extrusion liquid supply syringe 8 Artificial cerebrospinal fluid 9 Instruction manual 10 Needle body 11 Passage 12 Blocking section 13 Front end 13a Front end face 13b Side face 14 Outlet path 14a Circular cone section 14b Cylindrical section 14c Opening 15 Outlet port 15a Front end 16 Cylindrical section 17 Tip section 20 Connection section 21 Tube insertion hole 21a Tip side section 22 Body insertion hole 25 Tapered surface 26 Puncture needle attachment section 28 Syringe attachment section 31 Connection tube 34 Syringe attachment section 35 Attachment port 36 Switching valve 44 Syringe mounting portion 45 Mounting port 46 Second switching valve

Claims (27)

An administration tube used for administering a cell suspension containing cells or cell structures into a living body, the administration tube having a first end to which a puncture needle can be attached and a second end to which a syringe can be attached, the administration tube being capable of holding therein a total amount of the cell suspension to be administered in one treatment in order to reduce clogging of the cells or the cell structures, and the total amount of the cell suspension to be administered in the one treatment ;
An administration kit , wherein the administration tube is flexible . An administration tube used for administering a cell suspension containing cells or cell structures into a living body, the administration tube having a first end to which a puncture needle can be attached and a second end to which a syringe can be attached, the administration tube being capable of holding therein a total amount of the cell suspension to be administered in one treatment in order to reduce clogging of the cells or the cell structures, and the total amount of the cell suspension to be administered in the one treatment;
The administration kit, wherein the administration tube has an overall length greater than that of the syringe. An administration tube used for administering a cell suspension containing cells or cell structures into a living body, the administration tube having a first end to which a puncture needle can be attached and a second end to which a syringe can be attached, the administration tube being capable of holding therein a total amount of the cell suspension to be administered in one treatment in order to reduce clogging of the cells or the cell structures, and the total amount of the cell suspension to be administered in the one treatment;
The administration kit, wherein the administration tube has an inner diameter smaller than an inner diameter of the syringe. The administration kit according to claim 1 , further comprising a container that contains the cell suspension to be filled into the administration tube. 5. The administration kit according to claim 1, further comprising a cell suspension supply syringe that can be attached to the second end of the administration tube and is used to supply the cell suspension into the administration tube. a cell suspension supply syringe that can be attached to the second end of the administration tube and is used to supply the cell suspension into the administration tube;
4. The administration kit according to claim 1, wherein the cell suspension supply syringe is filled with the entire amount of the cell suspension. The administration kit according to claim 1 , wherein the entire amount of the cell suspension is filled in the administration tube. 7. The administration kit of claim 1, further comprising instructions for instructing a user to begin administration of the cell suspension after filling the administration tube with the entire amount of the cell suspension to be administered in one treatment. 9. The administration kit according to claim 8, wherein the instruction manual includes a first instruction to rotate the cell suspension supplying syringe back and forth multiple times around its central axis before starting to supply the cell suspension to the administration tube from the cell suspension supplying syringe filled with the entire amount of the cell suspension to be administered in the single treatment, and a second instruction to fill the entire amount of the cell suspension into the administration tube by starting to supply the cell suspension from the cell suspension supplying syringe to the administration tube after the cell suspension in the cell suspension supplying syringe is made uniformly dispersed. 10. The administration kit of claim 9 , wherein the instruction manual includes a third instruction to supply the entire amount of the cell suspension to the administration tube within 2 seconds after the cell suspension is uniformly dispersed. 11. The administration kit of claim 1, wherein the administration tube has a capacity of 2 mL or more. The administration kit according to claim 1 , wherein the administration tube has an inner diameter of 1.0 mm or more and 1.7 mm or less. The administration kit according to claim 1 , wherein the administration tube is up to 3 m in length. The administration kit according to claim 1 , wherein the administration tube has a capacity capable of accommodating the cell suspension that is greater than the total volume of the cell suspension to be administered in one treatment. The administration kit according to claim 1 , wherein the administration tube is provided with a puncture needle attachment portion for attaching the puncture needle to the first end. The administration kit of claim 1 , wherein the administration tube has a syringe attachment portion for attaching a syringe to the second end. The administration kit according to claim 16, wherein the syringe mounting portion includes a plurality of mounting ports to which the syringes can be attached, and a switching valve disposed between the administration tube and the mounting ports, for switching a communication state between the administration tube and the plurality of mounting ports. The administration kit of claim 1 , wherein the cell suspension contains the cell structures, and the cell structures have a diameter of 100 μm or more. The administration kit of claim 18 , wherein the diameter of the cell structure is 100 μm or more and 500 μm or less. The administration kit described in claim 18 or 19, wherein the cell structure is a structure comprising a biocompatible polymer block and at least one type of cell, and the plurality of polymer blocks are arranged in the gaps between the plurality of cells . 21. The administration kit of claim 1, further comprising an extrusion fluid supply syringe attachable to the second end of the administration tube and holding an extrusion fluid for extruding the cell suspension filled in the administration tube. The administration kit according to claim 21 , wherein the extrusion force required to extrude the extrusion liquid from the extrusion liquid supply syringe at an extrusion rate of 12 mL/min is 15 N or less. 23. The administration kit of claim 1, further comprising a puncture needle attached to the first end of the administration tube. The puncture needle includes a needle body along a front-rear direction, the needle body having a passage for the cell suspension formed therein, and an outlet for the cell suspension formed on an outer surface of a front end of the needle body,
the puncture needle includes a connection portion connected to the administration tube at the rear end of the needle body, the connection portion including a body insertion hole into which the rear end of the needle body is inserted, and a tube insertion hole communicating with the body insertion hole and into which the first end of the administration tube is inserted;
The administration kit according to claim 23, wherein the tip portion of the tube insertion hole connected to the main body insertion hole has a funnel shape whose inner diameter gradually narrows from the base end side to the tip end side, and the opening angle of the funnel shape opening from the tip end side to the base end side is 120° or less. The administration kit according to claim 24 , wherein the inner diameter of the passage of the needle body of the puncture needle is 0.5 mm or more. The administration kit according to claim 24 or 25 , wherein the inner diameter of the passage of the needle body of the puncture needle is at least twice the diameter of the cells or cell structures contained in the cell suspension. The administration kit according to any one of claims 24 to 26 , wherein the opening area of the outlet of the puncture needle is equal to or larger than the cross-sectional area of the passage of the needle body.