CN107213455A - A kind of stem cell medicine and its preparation method and application - Google Patents
A kind of stem cell medicine and its preparation method and application Download PDFInfo
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- CN107213455A CN107213455A CN201710442976.8A CN201710442976A CN107213455A CN 107213455 A CN107213455 A CN 107213455A CN 201710442976 A CN201710442976 A CN 201710442976A CN 107213455 A CN107213455 A CN 107213455A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/16—Blood plasma; Blood serum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1808—Epidermal growth factor [EGF] urogastrone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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Abstract
The application belongs to stem cells technology field, and in particular to a kind of stem cell medicine and its preparation method and application.A kind of stem cell medicine provided by the present invention, including：Fat mesenchymal stem cell, Platelet-rich plasm and EGF, each component compatibility is reasonable, Synergistic, jointly promotes the wound healing of diabetes；The present invention is using fat mesenchymal stem cell as seed cell, and collection is simple, be readily available, and immunogenicity is low, no ethics problem, significant for diabetic；Preparation method provided by the present invention, step operation is simple, process optimization, and large-scale production can be achieved.
Description
Technical field
The invention belongs to stem cells technology field, and in particular to a kind of stem cell medicine and its preparation method and application.
Background technology
With social progress and people's life, the transformation of diet style, diabetes morbidity increases year by year.China recent one
Epidemiological study result shows, Chinese diabetes prevalence surpasses the world average level 6.4% up to 9.7%.Glycosuria
One of foot disease is to cause diabetic disabled, or even dead severe chronic complication.With the hair of China human mortality aging
Exhibition, the diabetes incidence of disease increases year by year, deals with improperly and is possible to meeting amputation or death, and 85% amputation is due to ulcer.Sugar
Urine foot disease ulcer reduces quality of life in patients with diabetes, extended hospital stay, is brought to society and family huge
Financial burden, therefore overall merit need to be carried out to the preventing and treating of diabetic foot ulcer, prevented and treated early.
Clinically being used to treat the method for diabetes has drug therapy, reconstructing blood vessel and intervention operation, however, diabetes
Patient is got old and weak more, often with cardio-cerebrovascular diseases, it is impossible to which tolerance operation bridging etc. is stimulated, and involves vascular lesion small more
Artery, some patientss lack distal section artery efferent tract, do not possess the condition of row vascular bypass and PCI.Therefore, compel to be essential
Seek a kind of preparation that diabetic foot ulcer can be promoted to repair or method.
The content of the invention
In view of this, it is an object of the invention to provide a kind of stem cell medicine and its preparation method and application, for repairing
Multiple diabetic foot ulcer, reduces amputation rate, improves the quality of life of patient.
The concrete technical scheme of the present invention is as follows：
The invention provides a kind of stem cell medicine, including：Fat mesenchymal stem cell, Platelet-rich plasm and epidermis
Growth factor.
It is preferred that, the cell density of the fat mesenchymal stem cell is 1 × 105~10 × 105Individual/mL.
It is preferred that, the concentration of volume percent of the Platelet-rich plasm is 5%.
It is preferred that, the working concentration of the EGF is 5ng/mL.
It is preferred that, the fat mesenchymal stem cell is the third generation to the 5th fat subsitutes mescenchymal stem cell.
It is preferred that, the formulation of the stem cell medicine is ejection preparation.
It is preferred that, the injection dosage of the stem cell medicine is 3~8mL/ times.
Present invention also offers the preparation method of above-mentioned stem cell medicine, by fat mesenchymal stem cell, rich in blood platelet
Blood plasma and EGF mixing, obtain the stem cell medicine.
The stem cell medicine obtained present invention also offers above-mentioned stem cell medicine or above-mentioned preparation method is preparing reparation
Application in the medicine of diabetic foot ulcer.
A kind of stem cell medicine provided by the present invention, including：Fat mesenchymal stem cell, Platelet-rich plasm and table
Skin growth factor, each component compatibility is reasonable, Synergistic, jointly promotes the wound healing of diabetes；The present invention uses fat
Fat mescenchymal stem cell is as seed cell, and collection is simple, be readily available, and immunogenicity is low, no ethics problem, for sugar
Urinate patient significant；Preparation method provided by the present invention, step operation is simple, process optimization, and scale can be achieved
Metaplasia is produced.
Embodiment
The invention provides a kind of stem cell medicine extremely preparation method and application, help to reduce amputation rate, improve and suffer from
The quality of life of person.Stem cell medicine provided by the present invention includes：Fat mesenchymal stem cell, Platelet-rich plasm, table
Skin growth factor and solvent.
Further, the cell density of the fat mesenchymal stem cell is 1 × 105~10 × 105Individual/mL.
Further, the concentration of volume percent of the Platelet-rich plasm is 5%.
Further, the working concentration of the EGF is 5ng/mL.
Further, the fat mesenchymal stem cell is the third generation to the 5th fat subsitutes stem cell.
The present invention does not have special limit to the source of fat mesenchymal stem cell, Platelet-rich plasm and EGF
System, using well known to those skilled in the art, such as fat mesenchymal stem cell of commercial source, Platelet-rich plasm and
EGF, can also be prepared using customary preparation methods well known to those skilled in the art.
Further, also include in the stem cell medicine：Solvent；The solvent is preferably stem cell factor.
The present invention is not particularly limited to the source of the solvent, is given birth to using stem cell well-known to those skilled in the art
The long factor, such as commercial source.In some embodiments of the invention, solvent is the stem cell of our company Sai Silandai series
Growth factor, main component is human stem cell growth factor, deionized water, butanediol, hexylene glycol, propane diols, compounded amino
Acid, glucan, extract solution from aloe, polyglutamic acid and vitamin E etc..
Further, the formulation of the stem cell medicine is ejection preparation；
The injection dosage of the stem cell medicine is preferably 3~8mL/ times, more preferably 5mL/ times.
Present invention also offers the preparation method of above-mentioned stem cell medicine, by fat mesenchymal stem cell, rich in blood platelet
Blood plasma, EGF and solvent are well mixed, obtain the stem cell medicine.
The medicine of reparation diabetic foot ulcer is being prepared present invention also offers above-mentioned stem cell medicine or above-mentioned preparation method
Application in thing.
Technical scheme is clearly and completely described below in conjunction with the specific embodiment of the invention, it is clear that
Described embodiment is a part of embodiment of the invention, rather than whole embodiments.Those skilled in the art should manage
Solution, modifies to the specific embodiment of the present invention or some technical characteristics is replaced on an equal basis, without departing from the present invention
The spirit of technical scheme, all should cover in the scope of protection of the invention.
Platelet-rich plasm (PRP) used in following examples prepares suit and is purchased from the graceful biotechnology of Zhejiang Bao Er
Co., Ltd, solvent derives from Guangzhou Salia Cosmetic Technology Co., Ltd., and 654-2 is from plant of Solanaceae Anisodus tanguticus
A kind of belladonna alkaloids of Anisodus tanguticus (Maxim) Pascher root and seed kind separation, 654-2 parenteral solutions
Purchased from Sinopharm Group Rongsheng Pharmaceutical Co., Ltd..
Embodiment 1
1st, it is formulated
Third generation fat mesenchymal stem cell 1 × 105Individual/mL；
Platelet-rich plasm (PRP) 5vol%；
EGF (EGF) 5ng/mL；
Solvent is supplied.
2nd, prepare
(1) collect and take P3 fat subsitutes mescenchymal stem cells, suck culture medium, add digestive juice and digested, afterwards with suitable
Measure serum and terminate digestion, gently blow and beat into single cell suspension, then 1500rpm centrifugation 5min, abandon supernatant, cell is washed with 4 DEG C of PBS
It is resuspended and mixes after 3 times, adjustment cell density to 107Individual/mL.
(2) prepare suit with PRP and prepare PRP.
(3) in the centrifuge tube of 50mL specifications, successively add in PRP, EGF, solvent and step (1) and prepare by formula rate
P3 fat subsitutes mescenchymal stem cells, be sufficiently stirred for mix.
Embodiment 2
1st, it is formulated
Forth generation fat mesenchymal stem cell 5 × 105Individual/mL；
Platelet-rich plasm (PRP) 5vol%；
EGF (EGF) 5ng/mL；
Solvent is supplied.
2nd, prepare
(1) collect and take P3 fat subsitutes mescenchymal stem cells, suck culture medium, add digestive juice and digested, afterwards with suitable
Measure serum and terminate digestion, gently blow and beat into single cell suspension, then 1500rpm centrifugation 5min, abandon supernatant, cell is washed with 4 DEG C of PBS
It is resuspended and mixes after 3 times, adjustment cell density to 107Individual/mL.
(2) prepare suit with PRP and prepare PRP.
(3) in the centrifuge tube of 50mL specifications, successively add in PRP, EGF, solvent and step (1) and prepare by formula rate
P3 fat subsitutes mescenchymal stem cells, be sufficiently stirred for mix.
Embodiment 3
1st, it is formulated
5th fat subsitutes mescenchymal stem cell 1 × 106Individual/mL；
Platelet-rich plasm (PRP) 5vol%；
EGF (EGF) 5ng/mL；
Solvent is supplied.
2nd, prepare
(1) collect and take P3 fat subsitutes mescenchymal stem cells, suck culture medium, add digestive juice and digested, afterwards with suitable
Measure serum and terminate digestion, gently blow and beat into single cell suspension, then 1500rpm centrifugation 5min, abandon supernatant, cell is washed with 4 DEG C of PBS
It is resuspended and mixes after 3 times, adjustment cell density to 107Individual/mL.
(2) prepare suit with PRP and prepare PRP.
(3) in the centrifuge tube of 50mL specifications, successively add in PRP, EGF, solvent and step (1) and prepare by formula rate
P3 fat subsitutes mescenchymal stem cells, be sufficiently stirred for mix.
Embodiment 4
The stem cell medicine obtained to embodiment 1 into embodiment 3 carries out microorganism detection and endotoxin detection detection.Its
In, microorganism detection (bacterium, fungi) is detected according to the 4th general rule 1100 of version Pharmacopoeia of People's Republic of China in 2015；
Endotoxin detection is detected according to the 4th general rule 1143 of version Pharmacopoeia of People's Republic of China in 2015.
Testing result finds that biological agent is in bacterium, fungi and endotoxic detection made from embodiment 1-3, in the moon
Property, the up-to-standard of the stem cell medicine is illustrated, the reparation available for wound.
Embodiment 5
The diabetic that 80 courses of disease are 7-20 is randomly selected, wherein, man 37, female 43, by a variety of causes
Cause diabetes.Patient is randomly divided into 4 groups, one of which is control group, remaining 3 groups are experimental group.Control group use by
The mixed liquor of the U of gentamicin 8-16 ten thousand, insulin 4-8U, 654-2 parenteral solution 10mg and physiological saline composition carries out eddy current type punching
The surface of a wound is washed, is dried afterwards with sterile cotton balls, covers with sterile gauze, once a day, continues 4 weeks.Remaining 3 groups of experimental group is applying one
As therapeutic scheme while, embodiment 3-5 stem cell medicine is injected intravenously respectively, once, each 5mL continues for injection in every 7 days
4 weeks.
Table 1 is the prognosis situation of patient, and different degrees of improvement, wherein experimental group is presented in control group and 3 groups of experimental groups
Recovery from illness number and effective number be above control group, illustrate that stem cell medicine provided by the present invention can effectively facilitate diabetes
The healing of sufficient patient's ulcer wound surface.
It is evaluation index below：
Recovery from illness：4 Zhou Nei new granulation tissues are all long complete, and the surface of a wound heals completely；
It is effective：The surface of a wound reduces 75% in 4 weeks, and new granulation tissue largely grows, no inflammatory exudate；
Take a turn for the better：The surface of a wound is reduced in 4 weeks, and new granulation tissue part is grown, and inflammatory exudate is less；
It is invalid：Noninvasive reduction of area is small in 4 weeks, and no new granulation tissue, inflammatory exudate is more.
Table 1
P<0.05。
Claims (9)
1. a kind of stem cell medicine, it is characterised in that including：Fat mesenchymal stem cell, Platelet-rich plasm and epidermis life
The long factor.
2. stem cell medicine according to claim 1, it is characterised in that the cell density of the fat mesenchymal stem cell
For 1 × 105~10 × 105Individual/mL.
3. stem cell medicine according to claim 1, it is characterised in that the percent by volume of the Platelet-rich plasm
Concentration is 5%.
4. stem cell medicine according to claim 1, it is characterised in that the working concentration of the EGF is
5ng/mL。
5. stem cell medicine according to claim 1, it is characterised in that the fat mesenchymal stem cell be the third generation extremely
5th fat subsitutes mescenchymal stem cell.
6. stem cell medicine according to claim 1, it is characterised in that the formulation of the stem cell medicine is made for injection
Agent.
7. stem cell medicine according to claim 6, it is characterised in that the injection dosage of the stem cell medicine is 3~
8mL/ times.
8. the preparation method of stem cell medicine described in a kind of claim 1 to 7 any one, it is characterised in that will be filled between fat
Matter stem cell, Platelet-rich plasm and EGF mixing, obtain the stem cell medicine.
9. the stem cell that preparation method described in the stem cell medicine or claim 8 described in claim 1 to 7 any one is obtained
Preparation is preparing the application in repairing diabetic foot ulcer medicine.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710442976.8A CN107213455A (en) | 2017-06-13 | 2017-06-13 | A kind of stem cell medicine and its preparation method and application |
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| CN201710442976.8A CN107213455A (en) | 2017-06-13 | 2017-06-13 | A kind of stem cell medicine and its preparation method and application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108542915A (en) * | 2018-06-29 | 2018-09-18 | 胡葵葵 | A kind of drug and preparation method thereof promoting wound healing |
| CN110433283A (en) * | 2019-08-04 | 2019-11-12 | 丰泽康生物医药（深圳）有限公司 | A kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation is used for the drug and its preparation method and application of Corticosteroid addictive dermatitis |
Citations (1)
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| CN104357383A (en) * | 2014-10-11 | 2015-02-18 | 张炳强 | Preparation method of human adipose-derived MSCs (mesenchymal stem cells) and application of human adipose-derived mesenchymal stem cell in preparation of medicine for treating diseases |
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Patent Citations (1)
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| CN104357383A (en) * | 2014-10-11 | 2015-02-18 | 张炳强 | Preparation method of human adipose-derived MSCs (mesenchymal stem cells) and application of human adipose-derived mesenchymal stem cell in preparation of medicine for treating diseases |
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| 付小兵: "《中华战创伤学第10卷战创伤修复、再生与康复》", 30 June 2016, 郑州大学出版社 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108542915A (en) * | 2018-06-29 | 2018-09-18 | 胡葵葵 | A kind of drug and preparation method thereof promoting wound healing |
| CN110433283A (en) * | 2019-08-04 | 2019-11-12 | 丰泽康生物医药（深圳）有限公司 | A kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation is used for the drug and its preparation method and application of Corticosteroid addictive dermatitis |
| CN110433283B (en) * | 2019-08-04 | 2023-12-26 | 丰泽康生物医药（深圳）有限公司 | Medicine for treating glucocorticoid-dependent dermatitis prepared by combining peripheral blood multipotential cell active substance with platelet-rich plasma, and preparation method and application thereof |
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Application publication date: 20170929 |