CN116478817A - Self-gravity high-flux membrane chip - Google Patents

Abstract

The application discloses a dead weight high flux membrane type chip. The membrane chip includes a first layer, a porous membrane, and a second layer. The first layer has a first culture chamber; the second layer is laminated on the first layer along the gravity direction, at least one of the first layer and the second layer is provided with a second culture chamber, and the second culture chamber is positioned on one side of the porous membrane away from the first culture chamber in the gravity direction; the second layer is provided with at least one first sample injection pool and at least one second sample injection pool, and the first sample injection port of the first sample injection pool and the second sample injection port of the second sample injection pool are higher than the second culture chamber in the gravity direction. According to the method, fluid can correspondingly enter the first culture chamber and the second culture chamber in a self-gravity fluid driving mode and is attached to the corresponding surface of the porous membrane, so that dynamic cell culture is simulated, the culture flux is improved, convenience is provided for constructing a complex organ model, and the complexity of perfusion equipment and the construction cost of the organ model are reduced.

Description

Self-gravity high-flux membrane chip

Technical Field

The application relates to the technical field of cell culture, in particular to a self-gravity high-flux membrane chip.

Background

The membrane chip technology is a technology for in-vitro cell three-dimensional culture in a chip, and components such as cells, fluid, gas, extracellular microenvironment and the like in the chip are precisely controlled by constructing a micro-channel, a micro-reaction chamber and other functional components, so that human body micro-tissues and micro-organs with biological functionality are generated. The film chip technology has the advantages of integration, low consumption, high flux, high simulation degree, quick analysis and the like, and has wide application prospect in the fields of new medicine research and development, disease models, personalized medicine, aerospace medicine and the like.

The human body transports nutrients and gases to various parts of the body through blood vessels, and exchanges nutrients, gases and metabolites through organ-blood vessel interfaces. In order to simulate the structure in human body, porous membranes are used in the research of partial membrane chip technology to construct the interface, and vascular endothelial cells and cells corresponding to organs are respectively cultured at two sides of the membrane to simulate the cell arrangement and interface functions of the organ-blood vessel interface. However, due to the complexity of the human body structure, a high requirement is put on the complexity of the model that the membrane chip can build, and the fluid in the membrane chip is required to flow so as to better simulate the growth environment of the human body micro-tissues and micro-organs, so that the membrane chip has a complex structure, high cost for building the organ model, complex operation in the building process and low flux.

Disclosure of Invention

The embodiment of the application provides a self-gravity high-flux membrane type chip, which can solve the problem of complex structure of the membrane type chip.

The embodiment of the application provides a self-gravity high-flux membrane type chip, which comprises a first layer, a porous membrane and a second layer.

The first layer has a first space comprising a first culture chamber; the porous membrane is arranged corresponding to the first culture chamber; the second layer is stacked on the first layer along the gravity direction, at least one of the first layer and the second layer is provided with a second space, the second space comprises a second culture chamber, and the second culture chamber is positioned on one side of the porous membrane away from the first culture chamber in the gravity direction; the second layer is provided with at least one first sample injection pool and at least one second sample injection pool, one end of the first sample injection pool is communicated with the first culture chamber, the other end of the first sample injection pool is communicated with the surface of the second layer, which is away from the first layer, to form a first sample injection port, one end of the second sample injection pool is communicated with the second culture chamber, the other end of the second sample injection pool is communicated with the surface of the second layer, which is away from the first layer, to form a second sample injection port, and in the gravity direction, the first sample injection port and the second sample injection port are both higher than the second culture chamber.

In some exemplary embodiments, the first space further comprises a first flow channel in communication with the first sample inlet and the first culture chamber, the first flow channel and the first culture chamber being in a first plane perpendicular to a direction of gravity; the second space further comprises a second runner communicated with the second sample inlet and the second culture chamber, and the second runner and the second culture chamber are positioned on a second plane perpendicular to the gravity direction; the first plane and the second plane are parallel to the porous membrane and are respectively arranged on two opposite sides of the porous membrane in the gravity direction.

In some exemplary embodiments, the first space is a rotationally symmetrical space or an axisymmetrical space; and/or the second space is a rotationally symmetrical space or an axially symmetrical space.

In some exemplary embodiments, the first layer has a first interface surface connected to the second layer, the first layer has the second space, and the second culture chamber and the second flow channel open at the first interface surface.

In some exemplary embodiments, the first layer comprises a culture layer and a bottom plate stacked in a gravitational direction; the culture layer comprises a first butt joint surface connected with the second layer and a second butt joint surface deviating from the first butt joint surface; the first culture chamber and the first flow channel are arranged on the second butt joint surface, and the second butt joint surface is connected with the bottom plate.

In some exemplary embodiments, an edge region of the porous membrane is secured to at least one of the first layer and the second layer; or, the membrane chip further includes a support ring to which an edge region of the porous membrane is fixed, the support ring being mounted to at least one of the first layer and the second layer.

In some exemplary embodiments, a surface of the second layer facing away from the first layer is provided with a first aperture extending toward the first layer into communication with the second culture chamber.

In some exemplary embodiments, a surface of the second layer facing away from the first layer is provided with a plurality of evaporation channels; the second layer is provided with a plurality of first sample inlets and a plurality of second sample inlets; and one evaporation tank is arranged at least one position among two adjacent first sample inlets, two adjacent second sample inlets and two adjacent first sample inlets and second sample inlets.

In some exemplary embodiments, the shape of the first culture chamber is similar to the shape of the second culture chamber and is a circular hole, a regular polygon hole, or a regular polygon corner hole.

In some exemplary embodiments, the first space, the second space, the first sample inlet, the second sample inlet, and the porous membrane form a set of culture units, the membrane chip has a plurality of sets of culture units, and any two sets of culture units are spaced apart.

In a second aspect, the application also provides application of the membrane chip in biological model culture and drug analysis.

Based on the dead weight high flux membrane type chip of this application embodiment, have following beneficial effect at least:

by means of self gravity fluid driving, fluid can correspondingly enter the first culture chamber and the second culture chamber and adhere to the corresponding surfaces of the porous membranes, and reciprocating motion is facilitated under the driving of driving equipment, so that the corresponding fluid in the first culture chamber and the second culture chamber can be in flowing contact with cells adhering to the surfaces of the porous membranes, and dynamic cell culture is simulated. In addition, the membrane chip does not need to be provided with a structure for driving fluid to flow independently, so that the space occupied by the structure for conveying fluid to the first culture chamber and the second culture chamber is reduced, more groups of structural units for culturing cells can be arranged in a limited space, and the culture flux is improved. The membrane chip of the embodiment of the application provides convenience for constructing a complex organ model, is also beneficial to establishing a standard three-dimensional organ model culture mode, and reduces the complexity of perfusion equipment and the cost for constructing the organ model.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Fig. 1 is a schematic perspective view of a film chip according to an embodiment of the present disclosure;

FIG. 2 is a schematic diagram of an exploded structure of a membrane chip according to one embodiment of the present application;

FIG. 3 is a schematic perspective view of a membrane chip with a plurality of culture units according to an embodiment of the present application;

FIG. 4 is a schematic perspective view of a membrane chip with a first hole according to one embodiment of the present application;

FIG. 5 is a schematic cross-sectional view of a membrane chip having a first aperture according to one embodiment of the present application;

FIG. 6 is a schematic perspective view of a membrane chip with a first hole and a plurality of culture units according to an embodiment of the present application;

FIG. 7 is a schematic perspective view of a film chip with an evaporation tank according to an embodiment of the present disclosure;

FIG. 8 is a schematic cross-sectional view of a membrane chip with an evaporation tank according to one embodiment of the present application;

FIG. 9 is an exploded view of a membrane chip with evaporation tanks according to one embodiment of the present application;

fig. 10 is a schematic perspective view of a membrane chip with an evaporation tank and a plurality of culture units according to an embodiment of the present application.

Reference numerals:

10. a membrane chip; 11. a culturing unit;

100. a first layer; 110. a culture layer; 111. a first mating surface; 113. a step surface; 120. a bottom plate;

101. a first space; 1011. a first culture chamber; 1012. a first flow passage; 1013. a transfer flow passage;

200. a second layer; 201. a second space; 2011. a second culture chamber; 2012. a second flow passage; 202. a first sample inlet; 203. a second sample inlet; 204. a first hole; 205. an evaporation tank; 2021. a first sample introduction pool; 2031. a second sample injection pool;

300. a porous membrane;

400. and a support ring.

Detailed Description

In order to make the objects, technical solutions and advantages of the present application more apparent, the present application will be further described in detail with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the present application.

The inventor finds that the membrane type chip only comprising one side runner will limit the variety of the cultured organ model, is unfavorable for the establishment of the complex model, and the membrane type chip with the upper runner and the lower runner can meet the establishment requirement of the complex model. However, in the related art, the membrane chip with the upper and lower flow channels is driven by a peristaltic pump or an air pump in a fluid driving mode, the device is complex and expensive, the cost for constructing the organ model is high, the construction process is complex to operate, and the flux is low. In addition, whatever culture mode (for example, chip culture or equipment culture) cannot meet the requirement of high-throughput model construction, the requirement of large-scale screening of medicines is difficult to meet, and the application of the culture mode is severely limited. Based on this, the embodiment of the application provides a self-gravity high-flux membrane chip.

As shown in fig. 1 and 2, which are schematic structural views of a self-gravity high-flux membrane chip 10 according to an embodiment of the present application, the membrane chip 10 includes a first layer 100, a porous membrane 300, and a second layer 200.

The first layer 100 has a first space 101, and the first space 101 includes a first culture chamber 1011. The second layer 200 is stacked on one side of the first layer 100, and at least one of the first layer 100 and the second layer 200 has a second space 201, the second space 201 includes a second culture chamber 2011, and the second culture chamber 2011 corresponds to the first culture chamber 1011. The porous membrane 300 is installed to at least one of the first layer 100 and the second layer 200 and disposed corresponding to the first culturing chamber 1011, and the second culturing chamber 2011 is located at a side of the porous membrane 300 facing away from the first culturing chamber 1011, such that one surface of the porous membrane 300 can be in contact with a substance in the first culturing chamber 1011 and the other surface can be in contact with a substance in the second culturing chamber 2011. The porous membrane 300 covers the junction between the first culture chamber 1011 and the second culture chamber 2011, and prevents substances in the first culture chamber 1011 and the second culture chamber 2011 from being directly exchanged without passing through the porous membrane 300.

At least one first sample inlet 202 communicated with the first space 101 and at least one second sample inlet 203 communicated with the second space 201 are formed on the surface of the second layer 200, which is away from the first layer 100, so that fluid entering the first space 101 through the first sample inlet 202 can reach the first culture chamber 1011 to be in contact with the porous membrane 300, and fluid entering the second space 201 through the second sample inlet 203 can reach the second culture chamber 2011 to be in contact with the porous membrane 300. For example, one of the first and second sample inlets 202 and 203 may be filled with a cell suspension containing cells, the cell suspension enters a corresponding culture chamber, and the cells are adsorbed on a single side of the porous membrane 300 to perform cell culture in a corresponding mode; alternatively, cell suspensions containing different cells are respectively poured into the first sample inlet 202 and the second sample inlet 203, the cell suspensions enter corresponding culture chambers, and the cells are adsorbed on the surface of the porous membrane 300 so as to perform bilateral cell growth in corresponding modes.

The second layer 200 has at least one first sample inlet 2021, wherein one end of the first sample inlet 2021 is communicated with the first culture chamber 1011, and the other end forms a first sample inlet 202 on the surface of the second layer 200 facing away from the first layer 100. The second layer 200 has at least one second sample injection well 2031, wherein one end of the second sample injection well 2031 is communicated with the second culture chamber 2011, and the other end forms a second sample injection port 203 on the surface of the second layer 200 facing away from the first layer 100. The fluid injected from the first sample inlet 202 passes through the first sample tank 2021 and then enters the first culture chamber 1011, and the fluid injected from the second sample inlet 203 passes through the second sample tank 2031 and then enters the second culture chamber 2011. In the gravity direction G, the first sample introduction tank 2021 and the second sample introduction tank 2031 have depths that prevent fluid from splashing on the one hand, and reserve a space for storing fluid on the other hand, so that the first culture chamber 1011 and the second culture chamber 2011 can be filled with fluid to infiltrate the cells adsorbed on the surface of the porous membrane 300, respectively.

Specifically, in the embodiment of the present application, the second layer 200 is disposed on the first layer 100 along the gravity direction G, in which the second culture chamber 2011 is located above the porous membrane 300 and the first culture chamber 1011 is located below the porous membrane 300. In the gravity direction G, the first sample inlet 202 and the second sample inlet 203 are both higher than the second culture chamber 2011, so that the fluid buffered in the first sample tank 2021 and the fluid buffered in the second sample tank 2031 can be driven by gravity fluid respectively, so that the fluid entering the first space 101 can reach the first culture chamber 1011 to infiltrate one surface of the porous membrane 300 under the driving of gravity, and the fluid entering the second space 201 can reach the second culture chamber 2011 to infiltrate the other surface of the porous membrane 300 under the driving of gravity.

When the membrane chip 10 is used for cell culture, the membrane chip 10 is mounted on a driving device, and the reciprocating state of the membrane chip 10 is regulated and controlled by controlling the driving device, so that corresponding fluids in the first culture chamber 1011 and the second culture chamber 2011 can be in flowing contact with cells attached to the surface of the porous membrane 300, and cell dynamic culture is simulated. In addition, the membrane chip 10 is driven by gravity fluid, and no structure for driving fluid to flow is required to be separately arranged, so that the space occupied by the structures for conveying fluid to the first culture chamber 1011 and the second culture chamber 2011 can be reduced, more groups of structural units for culturing cells can be arranged in a limited space, and the culture flux can be improved. For example, as shown in fig. 3, the first space 101, the second space 201, the first sample inlet 202, the second sample inlet 203 and the porous membrane 300 form a group of culture units 11, the membrane chip 10 has a plurality of groups of culture units 11, any two groups of culture units 11 are arranged at intervals, the distance between two adjacent groups of culture units 11 can be designed to be closer, the space of the membrane chip 10 can be fully utilized, and the culture flux of the membrane chip 10 can be effectively improved.

The membrane chip 10 of the embodiment of the application provides convenience for constructing a complex organ model, is also beneficial to establishing a standard three-dimensional organ model culture mode, and can directly perfuse fluid from the first sample inlet 202 and the second sample inlet 203, so that the complexity of perfusion equipment and the cost for constructing the organ model are reduced.

Alternatively, the number of the first injection ports 202 communicating with the first culture chamber 1011 is plural; and/or the number of the second sample inlets 203 communicating with the second culture chamber 2011 is plural. By regulating the components of the fluid injected from each of the first sample inlet 202 and the second sample inlet 203, a rich organ model can be established, for example, the components of the drug contained in the injected fluid can be regulated, and the large-scale screening requirement of the drug can be satisfied.

To better simulate the cell dynamic environment, the first space 101 further comprises a first flow channel 1012, wherein one end of the first flow channel 1012 is directly communicated with the first culture chamber 1011, and the other end of the first flow channel 1012 is communicated with the first sample introduction pool 2021. The second space 201 further includes a second flow path 2012, wherein one end of the second flow path 2012 is directly communicated with the second culture chamber 2011, and the other end is communicated with the second sample injection well 2031. Alternatively, the flow area of the first flow channel 1012 is unchanged in the flow direction of the first flow channel 1012; alternatively, the flow area of the first flow channel 1012 gradually decreases in the flow direction toward the first culture chamber 1011 so as to regulate the flow rate or the flow velocity of the fluid entering the first culture chamber 1011, and the like. Alternatively, the first flow channel 1012 is a strip-shaped flow channel, and the extending direction of the first flow channel is designed so as to regulate the flow direction of the fluid entering the first culture chamber 1011. Likewise, the flow conditions such as the flow rate, the flow velocity, the flow direction, etc. of the fluid entering the second culture chamber 2011 through the second flow passage 2012 can be controlled by controlling the shape of the second flow passage 2012.

The first flow channel 1012 and the first culture chamber 1011 are positioned on a first plane perpendicular to the gravity direction G, the second flow channel 2012 and the second culture chamber 2011 are positioned on a second plane perpendicular to the gravity direction G, and the first plane and the second plane are parallel to the porous membrane 300 and are respectively arranged on two opposite sides of the porous membrane 300 in the gravity direction G, so that corresponding fluid can smoothly and stably enter and exit the first culture chamber 1011 and the second culture chamber 2011, the flow direction of the fluid is parallel to the flattened membrane surface of the porous membrane 300, and the fluid can more stably contact with cells adsorbed on the porous membrane 300, thereby being beneficial to establishing a standard three-dimensional organ model culture mode.

Optionally, when the number of the first sample inlets 202 is plural, the first space 101 includes plural first channels 1012 corresponding to the same number of the first channels 1012, so that the fluid in the first culture chamber 1011 can flow more smoothly through the cells adsorbed on the surface of the porous membrane 300, so as to better simulate the cell dynamic culture process. Further, the plurality of first flow passages 1012 in direct communication with the first culture chamber 1011 are arranged in rotational symmetry or in axial symmetry.

Optionally, when the number of the second sample inlets 203 is plural, the second space 201 includes plural second flow passages 2012 corresponding to the same number of the second flow passages 2012, so that the fluid in the second culture chamber 2011 can flow more smoothly through the cells adsorbed on the surface of the porous membrane 300, so as to better simulate the cell dynamic culture process. Further, the plurality of second flow passages 2012 are disposed rotationally symmetrically or axially symmetrically.

Further, the first space 101 is a rotationally symmetrical space or an axisymmetrical space, specifically, the first space 101 is a rotationally symmetrical space centered on the center of the first culture chamber 1011, or the first space 101 is an axisymmetrical space symmetrical with respect to a plane passing through the center of the first culture chamber 1011. Alternatively, the second space 201 is a rotationally symmetrical space or an axisymmetrical space, specifically, the second space 201 is a rotationally symmetrical space centered on the center of the second culture chamber 2011, or the second space 201 is an axisymmetrical space centered on a plane passing through the center of the second culture chamber 2011. In this way, the first space 101 and the second space 201 have a symmetrical structure, which is helpful for the corresponding fluid to flow more smoothly through the cells adsorbed on the surface of the porous membrane 300 when being reciprocally moved by the driving device. For example, when the driving device drives the organ chip to rotationally reciprocate in a preset direction, the first space 101 and the second space 201 are provided to be rotationally symmetrical spaces each; or, when the driving device drives the organ chip to reciprocate along a straight line, the first space 101 and the second space 201 are provided to be axisymmetric spaces, respectively.

In order to prevent the flow from splashing out of the first sample inlet 202 and the second sample inlet 203 during the movement of the organ chip, it is understood that the larger the distance from the first sample inlet 202 to the first culture chamber 1011 is, the better the splashing prevention effect is, and the larger the distance from the second sample inlet 203 to the second culture chamber 2011 is, the better the splashing prevention effect is. Optionally, the second layer 200 has a second space 201, specifically, a surface of the second layer 200 for connecting with the first layer 100 is provided with the second space 201, a surface of the first layer 100 for connecting with the second layer 200 is provided with the first space 101, and the porous membrane 300 is sandwiched between the first layer 100 and the second layer 200; alternatively, the first layer 100 has the second space 201, specifically, the first layer 100 has the first butt surface 111 for connecting with the second layer 200, the second culture chamber 2011 and the second flow passage 2012 are opened on the first butt surface 111, and the porous membrane 300 is provided on the first layer 100. By the arrangement, the second space 201 is located in a region far away from the first sample inlet 202 and the second sample inlet 203, and good splash-proof effect is achieved.

When the first layer 100 has the second space 201, the first space 101 further includes a transfer flow channel 1013 communicating with the first flow channel 1012, and the transfer flow channel 1013 extends to the first abutting surface 111 for communicating with the first sample cell 2021, so that when the first layer 100 and the second layer 200 are stacked, the first space 101 in the lower layer communicates with the first sample cell 2021. For example, the transfer flow path 1013 extends to the first junction 111 in the lamination direction of the first layer 100 and the second layer 200.

As shown in fig. 1 to 7, the first layer 100 includes a culture layer 110 and a bottom plate 120 stacked in a gravitational direction G; one of the surfaces of the culture layer 110 forms a first abutment surface 111, and the culture layer 110 further comprises a second abutment surface facing away from the first abutment surface 111, the second abutment surface being adapted to be coupled to the base plate 120. The first culturing chamber 1011 and the first flow channel 1012 are disposed on a second docking surface, and the second docking surface is hermetically connected to the bottom plate 120.

Optionally, when the first layer 100 has the first space 101 and the second space 201 at the same time, the second culture chamber 2011 and the second flow passage 2012 are opened on the first butt joint surface 111 of the culture layer 110, and the first culture chamber 1011 and the first flow passage 1012 are opened on the second butt joint surface of the culture layer 110, and by setting the first flow passage 1012 and the second flow passage 2012 on the surface of the culture layer 110, the processing and forming of the culture layer 110 are facilitated, and meanwhile, the first space 101 and the second space 201 have good sealing effect, so that the whole membrane chip 10 has a simple structure and is easy to process.

The second layer 200, the culture layer 110, the bottom plate 120 and the porous membrane 300 all have light transmittance so as to observe the growth state of cells adsorbed to the porous membrane 300. Alternatively, the second layer 200, the culture layer 110, and the bottom plate 120 may each be independently a glass plate, a plastic plate, a PDMS (polydimethylsiloxane) plate, etc., and the porous membrane 300 may be a plastic film such as a PC (Polycarbonate) film or a PET (polyethylene terephthalate ) film. The second layer 200, the culture layer 110 and the bottom plate 120 can be bonded and packaged by hot pressing, ultrasonic wave, laser, etc.

It is understood that the higher the mean one degree of the porous membrane 300, the more stable the growth state of the cells adsorbed on the surface of the porous membrane 300. Optionally, the edge area of the porous membrane 300 is fixed to at least one of the first layer 100 and the second layer 200, for example, the porous membrane 300 is bonded and encapsulated to the first layer 100 (specifically, the culture layer 110) by means of hot pressing, ultrasonic waves, laser, or the like; alternatively, as shown in fig. 8, the membrane chip 10 further includes a support ring 400, the edge region of the porous membrane 300 is fixed to the support ring 400, the support ring 400 is mounted on at least one of the first layer 100 and the second layer 200, for example, when the first layer 100 has the second space 201, the junction of the first culture chamber 1011 and the second culture chamber 2011 forms a step, the step has a step surface 113 perpendicular to the gravity direction G, and the support ring 400 is disposed on the step surface 113, thereby improving the uniform stability of the opening of the porous membrane 300.

As shown in fig. 4 and fig. 5, and fig. 7 and fig. 8, the surface of the second layer 200 facing away from the first layer 100 is provided with a first hole 204, and the first hole 204 extends toward the first layer 100 to be communicated with the second culture chamber 2011, so that the substance in the second culture chamber 2011 can directly pass through the first hole 204 to enter and exit the second culture chamber 2011. For example, taking a skin epithelial cell as an example, one of the methods of using the membrane chip 10 of the present embodiment is described as follows:

the cell suspension containing the skin epithelial cells was manually introduced into the second culture chamber 2011 above the porous membrane 300 through the first well 204 using a pipette. A pipette is used manually to inject the medium from at least one of the first injection ports 202 and fill the entire first culture chamber 1011. After the membrane chip 10 is placed in an incubator for stationary culture for a period of time, skin epithelial cells are deposited and adsorbed on the upper surface of the porous membrane 300. The cell suspension (the liquid remaining after the skin epithelial cells are adsorbed on the surface of the porous membrane 300) in the second culture chamber 2011 is sucked dry through the first hole 204 by using a pipette manually, so that the skin epithelial cells adsorbed on the upper surface of the porous membrane 300 are exposed to the air, and the growth environment of normal skin is simulated. The membrane chip 10 is placed on a shaking table, and the shaking table swings reciprocally through a specific angle, so that the culture medium in the first culture chamber 1011 flows back and forth among the plurality of first sample inlets 202, and the continuously reciprocally flowing culture medium forms dynamic culture of the skin epithelial cells through the porous membrane 300. The method can be used for researching the influence of the medicine on the skin epithelial cells by regulating and controlling the medicine components and the dosage contained in the culture medium.

In another embodiment, when cells need to grow on both sides of the single porous membrane 300, the cell suspension is injected into the first culture chamber 1011 through the first sample inlet 202, the cell suspension is injected into the second culture chamber 2011 through the second sample inlet 203, the membrane chip 10 is turned over each time after the cells on the corresponding surface of the porous membrane 300 are adsorbed and stabilized, the cell suspension (the liquid remaining after the cells are adsorbed on the surface of the porous membrane 300) is poured out, and the corresponding culture medium is injected into the first culture chamber 1011 and the second culture chamber 2011, so that the cell culture on both sides of the porous membrane 300 can be performed.

Since the membrane chip 10 needs to be moved frequently during the culturing process, evaporation of the culture medium is accelerated, the first holes 204 can be used to reserve space for storing more culture medium, and prevent the liquid in the second culture chamber 2011 from being evaporated.

Alternatively, as shown in fig. 7 to 10, a plurality of evaporation tanks 205 are provided on the surface of the second layer 200 facing away from the first layer 100, and the evaporation tanks 205 are spaced from the first space 101, the second space 201, the first sample inlet 202 and the second sample inlet 203. Each evaporation tank 205 is configured to store an evaporation preventing liquid, for example, the evaporation preventing liquid includes deionized water or PBS buffer. During cell culture, the evaporation preventing liquid in the evaporation tank 205 is exposed to the surface layer of the membrane chip 10, and the evaporation preventing liquid can be evaporated and take away heat, reducing the evaporation amount of the culture medium in both the first space 101 and the second space 201.

The second layer 200 has a plurality of first sample inlets 202 and a plurality of second sample inlets 203; an evaporation tank 205 is arranged at least one position among two adjacent first sample inlets 202, two adjacent second sample inlets 203 and two adjacent first sample inlets 202 and second sample inlets 203, so that the evaporation area is increased, and the positive drying prevention effect is improved.

The shape of the first culturing chamber 1011 is similar to that of the second culturing chamber 2011, and may be circular, regular polygon or regular polygonal rounded corners. Preferably, as shown in fig. 10, the first culture chamber 1011 and the second culture chamber 2011 are both circular, so that the shapes and positions of the first sample inlet 202, the second sample inlet 203, the first hole 204 and the evaporation tank 205 are convenient to be arranged, so that the four layouts occupy smaller area, more groups of culture units 11 are beneficial to being arranged in a limited space, and the flux of the membrane chip 10 is improved.

The same or similar reference numerals in the drawings of the present embodiment correspond to the same or similar components; in the description of the present application, it should be understood that, if there is an azimuth or positional relationship indicated by terms such as "upper", "lower", "left", "right", etc., based on the azimuth or positional relationship shown in the drawings, this is for convenience of description and simplification of the description, but does not indicate or imply that the apparatus or element to be referred must have a specific azimuth, be constructed and operated in a specific azimuth, and thus terms describing the positional relationship in the drawings are merely used for illustration and are not to be construed as limitations of the present patent, and that the specific meaning of the terms described above may be understood by those of ordinary skill in the art according to the specific circumstances.

The foregoing description of the preferred embodiments of the present application is not intended to be limiting, but is intended to cover any and all modifications, equivalents, and alternatives falling within the spirit and principles of the present application.

Claims (11)

1. A self-gravity high flux membrane chip, comprising:

a first layer having a first space comprising a first culture chamber;

a porous membrane disposed in correspondence with the first culture chamber; and

A second layer laminated on the first layer along the gravity direction, wherein at least one of the first layer and the second layer is provided with a second space, the second space comprises a second culture chamber, and the second culture chamber is positioned on one side of the porous membrane away from the first culture chamber along the gravity direction; the second layer is provided with at least one first sample injection pool and at least one second sample injection pool, one end of the first sample injection pool is communicated with the first culture chamber, the other end of the first sample injection pool is communicated with the surface of the second layer, which is away from the first layer, to form a first sample injection port, one end of the second sample injection pool is communicated with the second culture chamber, the other end of the second sample injection pool is communicated with the surface of the second layer, which is away from the first layer, to form a second sample injection port, and in the gravity direction, the first sample injection port and the second sample injection port are both higher than the second culture chamber.

2. The gravity high flux membrane chip according to claim 1, wherein,

the first space further comprises a first runner communicated with the first sample inlet and the first culture chamber, and the first runner and the first culture chamber are positioned on a first plane perpendicular to the gravity direction;

the second space further comprises a second runner communicated with the second sample inlet and the second culture chamber, and the second runner and the second culture chamber are positioned on a second plane perpendicular to the gravity direction;

the first plane and the second plane are parallel to the porous membrane and are respectively arranged on two opposite sides of the porous membrane in the gravity direction.

3. The gravity high-flux membrane chip according to claim 1 or 2, wherein,

the first space is a rotationally symmetrical space or an axially symmetrical space; and/or the number of the groups of groups,

the second space is a rotationally symmetrical space or an axially symmetrical space.

4. The gravity high-flux membrane chip of claim 2, wherein the first layer has a first butt surface connected to the second layer, the first layer has the second space, and the second culture chamber and the second flow channel are opened at the first butt surface.

5. The gravity-based high-flux membrane chip of claim 2, wherein the first layer comprises a culture layer and a bottom plate stacked in a gravitational direction; the culture layer comprises a first butt joint surface connected with the second layer and a second butt joint surface deviating from the first butt joint surface;

the first culture chamber and the first flow channel are arranged on the second butt joint surface, and the second butt joint surface is connected with the bottom plate.

6. The gravity high flux membrane chip according to claim 1, wherein,

an edge region of the porous membrane is fixed to at least one of the first layer and the second layer; or alternatively, the first and second heat exchangers may be,

the membrane chip further includes a support ring to which an edge region of the porous membrane is fixed, the support ring being mounted to at least one of the first layer and the second layer.

7. The self-gravity high-flux membrane chip of claim 1, wherein a first hole is provided in a surface of the second layer facing away from the first layer, the first hole extending toward the first layer to communicate with the second culture chamber.

8. The self-gravity high-flux membrane chip of claim 1 or 7, wherein a surface of the second layer facing away from the first layer is provided with a plurality of evaporation tanks;

the second layer is provided with a plurality of first sample inlets and a plurality of second sample inlets; and one evaporation tank is arranged at least one position among two adjacent first sample inlets, two adjacent second sample inlets and two adjacent first sample inlets and second sample inlets.

9. The gravity-based high-flux membrane chip of claim 1, wherein the first culture chamber has a shape similar to the shape of the second culture chamber and is a circular hole, a regular polygon hole, or a regular polygon hole.

10. The gravity high-flux membrane chip of any one of claims 1 to 9, wherein the first space, the second space, the first sample inlet, the second sample inlet and the porous membrane form a set of culture units, the membrane chip has a plurality of sets of the culture units, and any two sets of the culture units are arranged at intervals.

11. Use of the membrane chip of claim 10 in biological model culture and drug analysis.