CN110433283A - 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 - Google Patents
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 Download PDFInfo
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- CN110433283A CN110433283A CN201910733119.2A CN201910733119A CN110433283A CN 110433283 A CN110433283 A CN 110433283A CN 201910733119 A CN201910733119 A CN 201910733119A CN 110433283 A CN110433283 A CN 110433283A
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- 210000005259 peripheral blood Anatomy 0.000 title claims abstract description 43
- 239000011886 peripheral blood Substances 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 210000004623 platelet-rich plasma Anatomy 0.000 title claims abstract description 36
- 239000003814 drug Substances 0.000 title claims abstract description 29
- 201000004624 Dermatitis Diseases 0.000 title claims abstract description 26
- 229940079593 drug Drugs 0.000 title claims abstract description 24
- 239000003246 corticosteroid Substances 0.000 title claims abstract description 15
- 210000000130 stem cell Anatomy 0.000 claims abstract description 15
- 239000000411 inducer Substances 0.000 claims abstract description 11
- 239000002904 solvent Substances 0.000 claims abstract description 10
- 230000001737 promoting effect Effects 0.000 claims abstract description 3
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- 210000002381 plasma Anatomy 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 6
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- 239000001963 growth medium Substances 0.000 claims description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
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- 239000012467 final product Substances 0.000 claims description 3
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- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 4
- 206010033733 Papule Diseases 0.000 description 3
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- 108010024636 Glutathione Proteins 0.000 description 2
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- 229930182816 L-glutamine Natural products 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 206010043189 Telangiectasia Diseases 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 229940116977 epidermal growth factor Drugs 0.000 description 2
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- AGBQKNBQESQNJD-UHFFFAOYSA-M lipoate Chemical compound [O-]C(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-M 0.000 description 2
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- 230000003442 weekly effect Effects 0.000 description 2
- GDSOZVZXVXTJMI-SNAWJCMRSA-N (e)-1-methylbut-1-ene-1,2,4-tricarboxylic acid Chemical compound OC(=O)C(/C)=C(C(O)=O)\CCC(O)=O GDSOZVZXVXTJMI-SNAWJCMRSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- JDLSRXWHEBFHNC-UHFFFAOYSA-N Ufenamate Chemical compound CCCCOC(=O)C1=CC=CC=C1NC1=CC=CC(C(F)(F)F)=C1 JDLSRXWHEBFHNC-UHFFFAOYSA-N 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
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- 239000006143 cell culture medium Substances 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000000586 desensitisation Methods 0.000 description 1
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- 230000005713 exacerbation Effects 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
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- 230000003902 lesion Effects 0.000 description 1
- 230000006996 mental state Effects 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- SBNFWQZLDJGRLK-UHFFFAOYSA-N phenothrin Chemical compound CC1(C)C(C=C(C)C)C1C(=O)OCC1=CC=CC(OC=2C=CC=CC=2)=C1 SBNFWQZLDJGRLK-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
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Classifications
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
-
- 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
- A61K35/14—Blood; Artificial blood
- A61K35/16—Blood plasma; Blood serum
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- A—HUMAN NECESSITIES
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- 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
- 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
- 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
- A61K38/1825—Fibroblast growth factor [FGF]
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- 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
- A61K38/1858—Platelet-derived growth factor [PDGF]
- A61K38/1866—Vascular endothelial growth factor [VEGF]
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- 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
- A61K38/19—Cytokines; Lymphokines; Interferons
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- 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
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/21—Interferons [IFN]
- A61K38/217—IFN-gamma
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- 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
- A61K38/22—Hormones
- A61K38/30—Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
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Abstract
The present invention relates to stem cell medicine technical fields, in particular to 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, including peripheral blood multipotential cell, platelet rich plasma, inducer and solvent.Pass through the synergistic effect of three, it can control and mitigate the inflammatory reaction of skin, the outburst of inflammatory reaction is effectively controlled within the acute knock-on phase for deactivating hormone, simultaneously also containing ingredient the effect of promoting cutaneous immunisation and repair skin in product, skin can be helped to improve resistance, allergy repeatedly is avoided, state of gradually getting well.
Description
Technical field
The present invention relates to stem cell medicine technical field, in particular to a kind of peripheral blood multipotential cell active matter joint is rich
Thrombocyte plasma preparation is used for the drug and its preparation method and application of Corticosteroid addictive dermatitis.
Background technique
The clinical manifestation of Corticosteroid addictive dermatitis be face red spot, papule, flush, telangiectasis,
Dry, tight, itch, furfur etc., its treatment of the cause of disease is difficult, and recurrent exerbation, patient applies remission after hormone, after deactivating
Symptom recurrence is further continued for being thusly-formed vicious circle using hormone medicine, and dermatitis is increasingly heavier, and recurrence frequency is higher and higher,
To causing very big puzzlement on outpatients mental state.What the disease treated current more application is the substitution external application of non-hormonal anti-inflammatory drug, but
Curative effect is not still very satisfied.
Summary of the invention
In view of the above technical problems, the purpose of the present invention is to provide a kind of peripheral blood multipotential cell active matter joint is rich
Thrombocyte plasma preparation is used for the drug and its preparation method and application of Corticosteroid addictive dermatitis, life provided by the invention
Object preparation has preferable therapeutic effect to for Corticosteroid addictive dermatitis.
In order to solve the above technical problems, specific technical solution:
A kind of joint platelet rich plasma preparation of peripheral blood multipotential cell active matter is used for Corticodependence skin
Scorching drug, including peripheral blood multipotential cell, platelet rich plasma, inducer and solvent.
Wherein, the inducer includes following component: the epidermal growth factor of 50-300ng/mL, 10-200ng/mL
Insulin-like growth factor-i, the basic fibroblast growth factor of 1-100ng/mL, 1-100ng/mL blood vessel endothelium
Growth factor, the gamma interferon of 100-2000U/mL, the hematopoietin of 0.1-10U/mL, 50-200ug/mL it is anti-
Bad hematic acid.
The peripheral blood multipotential cell is no less than 200 × 10 in content4A cell/mL.In the platelet rich plasma
It is at least 300 × 10 containing concentration3The blood platelet of a cell/mL, red blood cell concentration are lower than 300 × 106A cell/mL.
The solvent is physiological saline.
The present invention also provides peripheral blood multipotential cell active matter joint platelet rich plasma preparations for sugared cortical hormone
The preparation method of the drug of plain dependence facial dermatitis, comprising the following steps: peripheral blood multipotential cell suspends be distributed in rich blood first
Mixture is obtained in platelet-poor plasma, then inducer and solvent are mixed to get solution, then mix mixture and solution, obtain
Peripheral blood multipotential cell active matter combines the drug that platelet rich plasma preparation is used for Corticosteroid addictive dermatitis.
Wherein, the peripheral blood multipotential cell is obtained by following steps:
(1) peripheral blood mononuclear cells trypsase 0.1-0.2g/mL and Collagenase 0.05-0.08g/mL, In
30-60min is digested at 37 DEG C, then 200 mesh are collected by filtration cell, then with erythrocyte cracked liquid, water-bath 3-5min under the conditions of 4 DEG C
Remove red blood cell to obtain the final product；
(2) peripheral blood mononuclear cells that step (1) obtains is seeded in stem cell media, in 37 DEG C, 5%CO2
Under the conditions of be incubated for 15-25h in the incubator, discard culture medium, rejoin in fresh stem cell media, connect to stem cell
When nearly 80-90% fusion, passage amplification cultivation is carried out；
(3) peripheral blood mononuclear cells that inoculation is in cell culture medium in step (2) is expanded into 5-10 generation, collected
Culture supernatant filters 0.22 μm of sterilizing filter of supernatant, merging filtrate.
Wherein the stem cell media is 1640 culture medium of RMPI, adds following component: cow's serum, 160-220ml/
L；PHA, 16-30g/L；L-Glutamine, 8-12g/L；Glutathione, 8-14g/L；Non sulphate glycosaminoglycan, 0.1-0.3g/
L；Lipoic acid, 0.1-0.3g/L；Penicillin, 0.005-0.01g/L.
The platelet rich plasma preparation method is aseptically, peripheral blood to be dispensed into PRP thixotroping separation gel and is mentioned
Pipe is taken, the 8-12min under 1500-2000g centrifugation, the blood plasma on thixotroping separation gel upper layer is platelet rich plasma.
The peripheral blood multipotential cell active matter combines platelet rich plasma preparation and is used for Corticosteroid addictive dermatitis
Drug application, preparation treatment be used for Corticosteroid addictive dermatitis drug application.
Compared with prior art, the present invention provides a kind of peripheral blood multipotential cell active matters to combine platelet rich plasma
Preparation is used for the drug and its preparation method and application of Corticosteroid addictive dermatitis, peripheral blood multipotential cell, Fu Xue little
Plate blood plasma, inducer can control and mitigate the inflammatory reaction of skin by the synergistic effect of three, in the urgency for deactivating hormone
Property the knock-on phase in effectively control the outburst of inflammatory reaction, while in product also containing promoting cutaneous immunisation and repairing skin the effect of
Ingredient can help skin to improve resistance, avoid allergy repeatedly, state of gradually getting well.
Specific embodiment
In order to make relevant technical staff in the field more fully understand technical solution of the present invention, below in conjunction with of the invention real
Mode is applied, the technical solution in embodiment of the present invention is clearly and completely described, it is clear that described embodiment
Only some embodiments of the invention, rather than whole embodiments.
A kind of joint platelet rich plasma preparation of peripheral blood multipotential cell active matter is used for Corticodependence skin
Scorching drug, including peripheral blood multipotential cell, platelet rich plasma, inducer and solvent.
Wherein, the inducer includes following component: the pancreas islet of the epidermal growth factor of 200ng/mL, 100ng/mL
Plain like growth factor -1, the basic fibroblast growth factor of 80ng/mL, 70ng/mL vascular endothelial growth factor,
The gamma interferon of 800U/mL, the hematopoietin of 5U/mL, 150ug/mL ascorbic acid.
The peripheral blood multipotential cell is no less than 200 × 10 in content4A cell/mL.In the platelet rich plasma
It is at least 300 × 10 containing concentration3The blood platelet of a cell/mL, red blood cell concentration are lower than 300 × 106A cell/mL.The solvent
For physiological saline.
The peripheral blood multipotential cell active matter combines platelet rich plasma preparation and is used for Corticosteroid addictive dermatitis
Drug preparation method, comprising the following steps: peripheral blood multipotential cell suspends first to be distributed in platelet rich plasma
To mixture, then inducer and solvent are mixed to get solution, then mix mixture and solution, obtain peripheral blood pluripotency
Cell activity Internet of Things close the drug that platelet rich plasma preparation is used for Corticosteroid addictive dermatitis.
Wherein, the peripheral blood multipotential cell is obtained by following steps: (1) peripheral blood mononuclear cells tryptose
Enzyme 0.2g/mL and Collagenase 0.05g/mL, digests 60min at 37 DEG C, and then 200 mesh are collected by filtration cell, then with red
Cell pyrolysis liquid, water-bath 5min removal red blood cell under the conditions of 4 DEG C to obtain the final product；(2) the single core of peripheral blood obtained step (1) is thin
Born of the same parents are seeded in stem cell media, in 37 DEG C, 5%CO2Under the conditions of be incubated in the incubator for 24 hours, discard culture medium, again
It is added in fresh stem cell media, when stem cell is merged close to 80-90%, carries out passage amplification cultivation；(3) by step
(2) peripheral blood mononuclear cells being seeded in stem cell media in expanded for 5 generations, collects culture supernatant, supernatant is used
0.22 μm of sterilizing filter filtering, merging filtrate.
Wherein the stem cell media is 1640 culture medium of RMPI, adds following component: cow's serum, 220ml/L；
PHA, 20g/L；L-Glutamine, 10g/L；Glutathione, 10g/L；Non sulphate glycosaminoglycan, 0.3g/L；Lipoic acid,
0.2g/L；Penicillin, 0.01g/L.
The platelet rich plasma preparation method is aseptically, peripheral blood to be dispensed into PRP thixotroping separation gel and is mentioned
Pipe is taken, the 12min under 2000g centrifugation, the blood plasma on thixotroping separation gel upper layer is platelet rich plasma.
For verification the verifying results, clinical test is taken:
1, patient is selected, treatment group and control group are randomly divided into.Treatment group and control group are in age, gender, the course of disease and disease
Shape scoring etc. is comparable without significant difference.Patient selection criteria has following (1), (2) item, and (3), (4)
At least one.
(1) there is the fixed position external application hormone product history of two weeks or more；(2) it deactivates containing 2~10 days originals after hormone product
There are disease or Repigmentation or exacerbation；There is hormone dependant phenomenon (to bounce, original disease or Repigmentation or add after being discontinued
Weight, symptom mitigates after reuse)；(3) typically skin lesion: using erythema, papule, drying and furfur as the more of basic lesions
Sample skin lesion, it is difficult to other skin diseases exponent；(4) subjective symptom: conscious part itch burns sample pain, tight swollen sense
Or it is dry uncomfortable, above-mentioned symptom heat aggravates, mitigates to the cold.
30 out-patients, male 5, female 25 are selected altogether；Age between 20~50 years old, external application hormone 2 months~3
Year, through multiple desensitization treatment, less effective.
2, treatment method: peripheral blood multipotential cell is no less than 200 × 10 in content4Treatment group: intracutaneous injection enters, note
Penetrate peripheral blood multipotential cell 2 × 106A mixed liquor, the next day additional 1 medication, dose is the same as the first time.1 treatment is observed weekly
Effect, 15 days are 1 course for the treatment of.
Control group: 5% Butyl flufenamate of external application observes weekly 1 curative effect 3 times a day, and 15 days are 1 treatment
Journey.
3, curative effect determinate standard
(1) skin lesion degree scores
4 objective indicators: flush, telangiectasis, pigment change and acneform eruptions；
1 subjective index: sense of discomfort.
Ranking criterion are as follows: 0 point be nothing, 1 point be it is slight, 2 points be moderate, 3 points be severe；Full marks are 15 points.
(2) clinical efficacy determines
The integrated value of patient is calculated separately before treating and after treatment, and calculates integral slip according to formula:
Integrate slip=(pre-treatment score-post treatment integral)/pre-treatment score × 100%.
By recovery from illness, effective, effective, invalid 4 standards, total effective rate is to fully recover plus effective calculating.It is discontinued after 1 week, occurs
Obvious flush, papule, itch etc. are recurrence.
Recovery from illness :≤1 is integrated after treatment.
It is effective: integral slip >=70%.
Effective: integral slip is 40%~69%.
Invalid: integral decreases below 40%.
4, result
Treatment group and control group complete the entire course for the treatment of as required respectively.Symptom score is shown in Table 1；Therapeutic effect is shown in Table 2.
1 treatment group of table is compared with control group symptom score
| Group | Number of cases | Before treatment | Treat 3d | Treat 7d | Treat 14d |
| Treatment group | 15 | 13.12±2.47 | 11.24±3.42 | 9.54±2.94 | 1.98±41.43 |
| Control group | 15 | 13.24±2.71 | 12.13±4.37 | 11.72±1.96 | 8.47±2.64 |
2 two groups of curative effects of table compare
| Group | It fully recovers (number of cases, %) | Effective (number of cases, %) | Effectively (number of cases, %) | (number of cases, %) in vain |
| Treatment group | 2471 | 1037 | 547 | 0 |
| Control group | 2415 | 954 | 8431 | 469 |
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation is used for Corticosteroid addictive dermatitis
Drug, which is characterized in that including peripheral blood multipotential cell, platelet rich plasma, inducer and solvent.
2. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 1 is for sugar
The drug of Corticodependence dermatitis, which is characterized in that the inducer includes following component: the table of 50-300ng/mL
Skin growth factor, the insulin-like growth factor-i of 10-200ng/mL, 1-100ng/mL basic fibroblast growth because
The promoting erythrocyte of son, the vascular endothelial growth factor of 1-100ng/mL, the gamma interferon of 100-2000U/mL, 0.1-10U/mL
Generate the ascorbic acid of element, 50-200ug/mL.
3. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 1 is for sugar
The drug of Corticodependence dermatitis, which is characterized in that the peripheral blood multipotential cell is no less than 200 × 10 in content4
A cell/mL.
4. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 1 is for sugar
The drug of Corticodependence dermatitis, which is characterized in that be at least 300 × 10 containing concentration in the platelet rich plasma3It is a
Cell/mL blood platelet, red blood cell concentration are lower than 300 × 106A cell/mL.
5. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 1 is for sugar
The drug of Corticodependence dermatitis, which is characterized in that the solvent is physiological saline.
6. being used according to claim 1 to a kind of joint platelet rich plasma preparation of peripheral blood multipotential cell active matter described in 5
In the preparation method of the drug of Corticosteroid addictive dermatitis, which comprises the following steps: peripheral blood pluripotency is thin
Born of the same parents suspend to be distributed in platelet rich plasma first obtains mixture, and then inducer and solvent are mixed to get solution, then will
Mixture and solution mixing obtain the joint platelet rich plasma preparation of peripheral blood multipotential cell active matter for glucocorticoid
The drug of dependence facial dermatitis.
7. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 6 is for sugar
The process for preparing medicine of Corticodependence dermatitis, which is characterized in that the peripheral blood multipotential cell passes through following steps
It obtains:
(1) peripheral blood mononuclear cells trypsase 0.1-0.2g/mL and Collagenase 0.05-0.08g/mL, at 37 DEG C
Lower digestion 30-60min, then 200 mesh are collected by filtration cell, then with erythrocyte cracked liquid, and water-bath 3-5min is removed under the conditions of 4 DEG C
Red blood cell to obtain the final product；
(2) peripheral blood mononuclear cells that step (1) obtains is seeded in stem cell media, in 37 DEG C, 5%CO2Under the conditions of
It is incubated for 15-25h in the incubator, discards culture medium, rejoins in fresh stem cell media, to stem cell close to 80-
When 90% fusion, passage amplification cultivation is carried out；
(3) peripheral blood mononuclear cells being seeded in stem cell media in step (2) is expanded into 5-10 generation, collected in culture
Clear liquid filters 0.22 μm of sterilizing filter of supernatant, merging filtrate.
8. a kind of peripheral blood multipotential cell active matter joint platelet rich plasma preparation according to claim 6 is for sugar
The process for preparing medicine of Corticodependence dermatitis, which is characterized in that the platelet rich plasma preparation method is, sterile
Under the conditions of, peripheral blood is dispensed into PRP thixotroping separation gel extraction tube, the 8-12min under 1500-2000g centrifugation, thixotroping separation gel
The blood plasma on upper layer is platelet rich plasma.
9. being used according to claim 1 to a kind of joint platelet rich plasma preparation of peripheral blood multipotential cell active matter described in 5
In the application of the drug of Corticosteroid addictive dermatitis, which is characterized in that be used for Corticodependence in preparation treatment
The application of dermatitis drug.
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