CN112245454A - 一种细胞疤痕修复方法 - Google Patents
一种细胞疤痕修复方法 Download PDFInfo
- Publication number
- CN112245454A CN112245454A CN202011110667.9A CN202011110667A CN112245454A CN 112245454 A CN112245454 A CN 112245454A CN 202011110667 A CN202011110667 A CN 202011110667A CN 112245454 A CN112245454 A CN 112245454A
- Authority
- CN
- China
- Prior art keywords
- stem cells
- culture
- culture solution
- mesenchymal stem
- adipose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 231100000241 scar Toxicity 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 22
- 210000000130 stem cell Anatomy 0.000 claims abstract description 39
- 210000003780 hair follicle Anatomy 0.000 claims abstract description 35
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims abstract description 31
- 210000004027 cell Anatomy 0.000 claims abstract description 17
- 238000012258 culturing Methods 0.000 claims abstract description 14
- 230000000694 effects Effects 0.000 claims abstract description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 238000004113 cell culture Methods 0.000 claims abstract description 11
- 230000003325 follicular Effects 0.000 claims abstract description 11
- 239000006228 supernatant Substances 0.000 claims abstract description 8
- 210000000577 adipose tissue Anatomy 0.000 claims abstract description 7
- 239000002609 medium Substances 0.000 claims abstract description 7
- 230000004069 differentiation Effects 0.000 claims abstract description 6
- 238000005516 engineering process Methods 0.000 claims abstract description 6
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 230000035755 proliferation Effects 0.000 claims abstract description 5
- 230000003321 amplification Effects 0.000 claims abstract description 4
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 4
- 238000000746 purification Methods 0.000 claims abstract description 4
- 229940090044 injection Drugs 0.000 claims description 34
- 239000007924 injection Substances 0.000 claims description 34
- 238000002347 injection Methods 0.000 claims description 34
- 239000000243 solution Substances 0.000 claims description 34
- 210000003491 skin Anatomy 0.000 claims description 21
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 20
- 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 20
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 10
- 229930003268 Vitamin C Natural products 0.000 claims description 10
- 229930003427 Vitamin E Natural products 0.000 claims description 10
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 10
- 230000008439 repair process Effects 0.000 claims description 10
- 235000019154 vitamin C Nutrition 0.000 claims description 10
- 239000011718 vitamin C Substances 0.000 claims description 10
- 235000019165 vitamin E Nutrition 0.000 claims description 10
- 229940046009 vitamin E Drugs 0.000 claims description 10
- 239000011709 vitamin E Substances 0.000 claims description 10
- 230000001413 cellular effect Effects 0.000 claims description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 229940093181 glucose injection Drugs 0.000 claims description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 3
- 239000004378 Glycyrrhizin Substances 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 239000012228 culture supernatant Substances 0.000 claims description 3
- 229960002344 dexamethasone sodium phosphate Drugs 0.000 claims description 3
- PLCQGRYPOISRTQ-FCJDYXGNSA-L dexamethasone sodium phosphate Chemical compound [Na+].[Na+].C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)COP([O-])([O-])=O)(O)[C@@]1(C)C[C@@H]2O PLCQGRYPOISRTQ-FCJDYXGNSA-L 0.000 claims description 3
- 239000012091 fetal bovine serum Substances 0.000 claims description 3
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 claims description 3
- 229960004949 glycyrrhizic acid Drugs 0.000 claims description 3
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 claims description 3
- 235000019410 glycyrrhizin Nutrition 0.000 claims description 3
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000002504 physiological saline solution Substances 0.000 claims description 3
- 210000002966 serum Anatomy 0.000 claims description 3
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 2
- 102000009024 Epidermal Growth Factor Human genes 0.000 claims 1
- 101800003838 Epidermal growth factor Proteins 0.000 claims 1
- 229940116977 epidermal growth factor Drugs 0.000 claims 1
- 206010039580 Scar Diseases 0.000 description 24
- 208000032544 Cicatrix Diseases 0.000 description 13
- 230000037387 scars Effects 0.000 description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 230000003203 everyday effect Effects 0.000 description 5
- 206010072170 Skin wound Diseases 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000019522 cellular metabolic process Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 210000001732 sebaceous gland Anatomy 0.000 description 3
- 102000018386 EGF Family of Proteins Human genes 0.000 description 2
- 108010066486 EGF Family of Proteins Proteins 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000036770 blood supply Effects 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 238000005562 fading Methods 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- 208000028990 Skin injury Diseases 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 210000001339 epidermal cell Anatomy 0.000 description 1
- 210000000442 hair follicle cell Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000037380 skin damage Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000037314 wound repair Effects 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
- A61K31/355—Tocopherols, e.g. vitamin E
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- 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
- A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
-
- 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
-
- 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
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
- C12N5/0627—Hair cells
- C12N5/0628—Hair stem cells; Hair progenitors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/38—Vitamins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/39—Steroid hormones
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/999—Small molecules not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Developmental Biology & Embryology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Dermatology (AREA)
- Immunology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Gastroenterology & Hepatology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Rheumatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种细胞疤痕修复方法,包括如下步骤:S1:采用FUE技术提取一个毛囊单位;S2:在电子显微镜下将所述毛囊单位分离出完整的单株毛囊;S3:将提取分离的单株毛囊放入特制培养液培养备用,进行培育达到增强毛囊干细胞活性,促进其分化增殖;S4:从自体分离脂肪组织并从中脂肪间充质干细胞;S5:将获得的脂肪间充质干细胞置于第一培养皿中进行纯化和扩增同时在第一培养基内加入混合基;S6:待S5中脂肪间充质干细胞扩增至70‑90%后去除上清物,往第一培养皿中加入无酚红培养基,培养48‑96小时,收取培养液,将培养液离心后,取上清液得间充质干细胞培养液;S7:将S6中获取的间充质干细胞培养液置于第二培养皿内。
Description
技术领域
本发明涉及细胞疤痕修复技术领域,具体为一种细胞疤痕修复方法。
背景技术
由于烫伤、擦伤、皮肤溃烂和烧伤等原因,可能会导致皮肤的大范围伤害;皮肤的损伤容易造成细菌感染,体液流失并引起各种临床并发症;皮肤损伤修复之后可能会存在疤痕影响皮肤美观,目前市售存在多种伤口修复产品,但大多不同程度地存在着化学成分过多、见效慢、产品价格昂贵等诸多不足。
发明内容
本发明的目的在于提供一种细胞疤痕修复方法,以解决上述背景技术中提出的问题。
为实现上述目的,本发明提供如下技术方案:
一种细胞疤痕修复方法,包括如下步骤:
S1:采用FUE技术从自体上提取一个毛囊单位;
S2:在电子显微镜下将所述毛囊单位分离出完整的单株毛囊;
S3:将提取分离的单株毛囊放入特制培养液培养备用,进行培育达到增强毛囊干细胞活性,促进其分化增殖;
S4:从自体上分离脂肪组织并从脂肪组织中提取脂肪间充质干细胞同时从自体上获取表皮因子;
S5:将获得的脂肪间充质干细胞置于第一培养皿中进行纯化和扩增,同时在第一培养基内加入混合基;
S6:待S5中脂肪间充质干细胞扩增至70-90%后去除上清物,往第一培养皿中加入无酚红培养基,培养48-96小时,收取培养液,将培养液离心后,取上清液得间充质干细胞培养液;
S7:将S6中获取的间充质干细胞培养液置于第二培养皿内,待S5中的脂肪间充质干细胞浓度达到106-108mg/ml时,将脂肪间充质干细胞以及S3中的毛囊干细胞转移至第二培养皿内,同时向第二培养皿内加入细胞载体、表皮因子、维生素C和维生素E,混合形成干细胞注射物。
作为本发明一种优选的技术方案,所述步骤S2中,提取毛囊单位放在在电子显微镜下分离出单株毛囊,在放大2-10倍的状态下进行分离,并尽可能的保留了提取全部皮肤、皮肤附属器和毛囊干细胞的完备性。
作为本发明一种优选的技术方案,所述步骤S3中特制培养液的温度为0-4℃,培养时间60分钟以上,培养液的主要成分为10%的葡萄糖注射液、复方甘草酸苷注射液和***磷酸钠注射液。
作为本发明一种优选的技术方案,所述步骤S5中的混合培养基包括DMEM/F12培养基和胎牛血清。
作为本发明一种优选的技术方案,所述维生素C的质量浓度为5-50mg/ml;维生素E的质量浓度为5-50mg/ml。
作为本发明一种优选的技术方案,所述步骤S7中的细胞载体为自体血清、0.9%生理盐水或5%葡萄糖注射液。
作为本发明一种优选的技术方案,表皮细胞生长因子(EGF)的质量浓度为10~50ng/mL,优选为15~45ng/mL,更优选为20~40ng/mL。
作为本发明一种优选的技术方案,步骤S6中培养液的离心力优选为200~400g,更优选为250~350g,最优选为300~320g;所述培养液的离心时间优选为5~10min,更优选为6~9min,最优选为7~8min;离心后取得的培养液上清,优选过0.22um的滤膜。
与现有技术相比,本发明的有益效果是:本发明提供的细胞疤痕修复方法,通过毛囊体外培养技术,放入特殊培养液培养,进行培育提高毛囊干细胞存活率,毛囊干细胞具有多向分化潜能,它可以分化成表皮、毛囊、皮脂腺,参与皮肤创伤愈合的过程,进而达到改善疤痕血供和软化疤痕的作用。
本发明采用脂肪间充质干细胞培养液,获取简单,且免疫原性低,对人体无害,并且脂肪间充质干细胞在培养过程中会分泌活性物质到培养液中,配合维生素C、维生素E和表皮细胞生长因子,能够有效抑制疤痕处色素的沉积,滋养肌肤,促进细胞的新陈代谢,从而达到淡化疤痕,修复皮肤创面的功效。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
具体的:本发明提供一种技术方案:一种细胞疤痕修复方法,包括如下步骤:
S1:采用FUE技术从自体上提取一个毛囊单位;
S2:在电子显微镜下将所述毛囊单位分离出完整的单株毛囊;
S3:将提取分离的单株毛囊放入特制培养液培养备用,进行培育达到增强毛囊干细胞活性,促进其分化增殖;
S4:从自体上分离脂肪组织并从脂肪组织中提取脂肪间充质干细胞同时从自体上获取表皮因子;
S5:将获得的脂肪间充质干细胞置于第一培养皿中进行纯化和扩增,同时在第一培养基内加入混合基;
S6:待S5中脂肪间充质干细胞扩增至70-90%后去除上清物,往第一培养皿中加入无酚红培养基,培养48-96小时,收取培养液,将培养液离心后,取上清液得间充质干细胞培养液;
S7:将S6中获取的间充质干细胞培养液置于第二培养皿内,待S5中的脂肪间充质干细胞浓度达到106-108mg/ml时,将脂肪间充质干细胞以及S3中的毛囊干细胞转移至第二培养皿内,同时向第二培养皿内加入细胞载体、表皮因子、维生素C和维生素E,混合形成干细胞注射物。
进一步的,所述步骤S2中,提取毛囊单位放在在电子显微镜下分离出单株毛囊,在放大2-10倍的状态下进行分离,并尽可能的保留了提取全部皮肤、皮肤附属器和毛囊干细胞的完备性。
进一步的,所述步骤S3中特制培养液的温度为0-4℃,培养时间60分钟以上,培养液的主要成分为10%的葡萄糖注射液、复方甘草酸苷注射液和***磷酸钠注射液。
进一步的,所述步骤S5中的混合培养基包括DMEM/F12培养基和胎牛血清。
进一步的,维生素C的质量浓度为5-50mg/ml;维生素E的质量浓度为5-50mg/ml;维生素C和维生素E可以抑制减少疤痕处色素的沉淀、滋养皮肤、促进细胞增殖代谢。
进一步的,所述步骤S7中的细胞载体为自体血清、0.9%生理盐水或5%葡萄糖注射液。
进一步的,所述维生素C和维生素E的质量浓度均为5~50mg/mL,优选为10~45mg/mL,更优选为15~40mg/mL,具体的,在本发明的实施例中,可以是5mg/mL,20mg/mL,或50mg/mL。
进一步的,表皮细胞生长因子(EGF)的质量浓度为10~50ng/mL,优选为15~45ng/mL,更优选为20~40ng/mL。
进一步的,步骤S6中培养液的离心力优选为200~400g,更优选为250~350g,最优选为300~320g;所述培养液的离心时间优选为5~10min,更优选为6~9min,最优选为7~8min;离心后取得的培养液上清,优选过0.22um的滤膜。
进一步的,招募存在痘疤的志愿者120人,随机分为6组,每组20人;对照组设有一组,实验组设有五组;对照组以生理盐水代替注射物,每天注射两针;实验组一以干细胞注射物(该注射物中脂肪间充质质量浓度为106mg/ml,干细胞培养了48小时)作为注射物,每天注射两针,连续注射一个月,对照组志愿者的痘疤并无改善,实验组存在6人痘疤完全消失即完全治愈,9人存在痘疤位置平整肤色自然即有明显效果,3人存在痘疤肉眼可辨的减轻即有效,2人与使用前无任何区别即无效;由上述实验数据可知,有效率为90%,治愈率为30%。
实验组二,以干细胞注射物(该注射物中脂肪间充质质量浓度为107mg/ml,干细胞培养了48小时)作为注射物,对照组以生理盐水代替注射物,同样每天注射两针,连续注射一个月,可以发现实验组存在8人痘疤完全消失即完全治愈,7人存在痘疤位置平整肤色自然即有明显效果,5人存在痘疤肉眼可辨的减轻即有效,0人与使用前无任何区别即无效;由上述实验数据可知,有效率为100%,治愈率为40%。
实验组三,以干细胞注射物(该注射物中脂肪间充质质量浓度为108mg/ml,干细胞培养了48小时)作为注射物,对照组以生理盐水代替注射物,同样每天注射两针,连续注射一个月,可以发现实验组存在9人痘疤完全消失即完全治愈,9人存在痘疤位置平整肤色自然即有明显效果,2人存在痘疤肉眼可辨的减轻即有效,0人与使用前无任何区别即无效;由上述实验数据可知,有效率为100%,治愈率为45%。
综上所述,由实验组1-3可知,在干细胞培养时间不变的情况下,干细胞浓度越高,该干细胞注射物的疤痕治愈率越高,即干细胞浓度最优选为108mg/ml。
实验组四,以干细胞注射物(该注射物中脂肪间充质质量浓度为106mg/ml,干细胞培养了72小时)作为注射物,对照组以生理盐水代替注射物,同样每天注射两针,连续注射一个月,可以发现实验组存在12人痘疤完全消失即完全治愈,8人存在痘疤位置平整肤色自然即有明显效果,0人存在痘疤肉眼可辨的减轻即有效,0人与使用前无任何区别即无效;由上述实验数据可知,有效率为100%,治愈率为60%。
实验组五,以干细胞注射物(该注射物中脂肪间充质质量浓度为106mg/ml,干细胞培养了96小时)作为注射物,对照组以生理盐水代替注射物,同样每天注射两针,连续注射一个月,可以发现实验组存在8人痘疤完全消失即完全治愈,6人存在痘疤位置平整肤色自然即有明显效果,4人存在痘疤肉眼可辨的减轻即有效,2人与使用前无任何区别即无效;由上述实验数据可知,有效率为90%,治愈率为40%。
综上所述,由实验组一、四和五可知,在干细胞浓度不变的情况下,脂肪间充质干细胞在无酚红培养基中的培养时间最优选为60~72小时,其余时间段为次优选。
在本发明中,本发明提供的细胞疤痕修复方法中,步骤简单明了易于操作,通过毛囊体外培养技术,放入特殊培养液培养,进行培育提高毛囊干细胞存活率,毛囊干细胞体外培养具有慢周期性、自我更新能力强、高增殖能力、多分化潜能等特性;根据所处环境不同,毛囊干细胞可分化为毛囊细胞、皮脂腺细胞、毛囊间表皮细胞等,这些细胞对于皮肤的重构和再生具有重要意义,因此可成功应用于疤痕的修复,治疗各种疤痕;毛囊干细胞具有多向分化潜能,它可以分化成表皮、毛囊、皮脂腺,参与皮肤创伤愈合的过程,进而达到改善疤痕血供和软化疤痕的作用。
本发明采用脂肪间充质干细胞培养液,获取简单,且免疫原性低,对人体无害,并且脂肪间充质干细胞在培养过程中会分泌活性物质到培养液中,配合维生素C、维生素E和表皮细胞生长因子,能够有效抑制疤痕处色素的沉积,滋养肌肤,促进细胞的新陈代谢,从而达到淡化疤痕,修复皮肤创面的功效。
尽管已经示出和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。
Claims (8)
1.一种细胞疤痕修复方法,其特征在于:包括如下步骤:
S1:采用FUE技术从自体上提取一个毛囊单位;
S2:在电子显微镜下将所述毛囊单位分离出完整的单株毛囊;
S3:将提取分离的单株毛囊放入特制培养液培养备用,进行培育达到增强毛囊干细胞活性,促进其分化增殖;
S4:从自体上分离脂肪组织并从脂肪组织中提取脂肪间充质干细胞同时从自体上获取表皮因子;
S5:将获得的脂肪间充质干细胞置于第一培养皿中进行纯化和扩增,同时在第一培养基内加入混合基;
S6:待S5中脂肪间充质干细胞扩增至70-90%后去除上清物,往第一培养皿中加入无酚红培养基,培养48-96小时,收取培养液,将培养液离心后,取上清液得间充质干细胞培养液;
S7:将S6中获取的间充质干细胞培养液置于第二培养皿内,待S5中的脂肪间充质干细胞浓度达到106-108mg/ml时,将脂肪间充质干细胞以及S3中的毛囊干细胞转移至第二培养皿内,同时向第二培养皿内加入细胞载体、表皮因子、维生素C和维生素E,混合形成干细胞注射物。
2.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:所述步骤S2中,提取毛囊单位放在在电子显微镜下分离出单株毛囊,在放大2-10倍的状态下进行分离,并尽可能的保留了提取全部皮肤、皮肤附属器和毛囊干细胞的完备性。
3.根据权利要求2所述的一种细胞疤痕修复方法,其特征在于:所述步骤S3中特制培养液的温度为0-4℃,培养时间60分钟以上,培养液的主要成分为10%的葡萄糖注射液、复方甘草酸苷注射液和***磷酸钠注射液。
4.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:所述步骤S5中的混合培养基包括DMEM/F12培养基和胎牛血清。
5.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:所述维生素C的质量浓度为5-50mg/ml;维生素E的质量浓度为5-50mg/ml。
6.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:所述步骤S7中的细胞载体为自体血清、0.9%生理盐水或5%葡萄糖注射液。
7.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:表皮细胞生长因子的质量浓度为10~50ng/mL,优选为15~45ng/mL,更优选为20~40ng/mL。
8.根据权利要求1所述的一种细胞疤痕修复方法,其特征在于:步骤S6中培养液的离心力优选为200~400g,更优选为250~350g,最优选为300~320g;所述培养液的离心时间优选为5~10min,更优选为6~9min,最优选为7~8min;离心后取得的培养液上清,优选过0.22um的滤膜。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011110667.9A CN112245454A (zh) | 2020-10-16 | 2020-10-16 | 一种细胞疤痕修复方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011110667.9A CN112245454A (zh) | 2020-10-16 | 2020-10-16 | 一种细胞疤痕修复方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN112245454A true CN112245454A (zh) | 2021-01-22 |
Family
ID=74244578
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011110667.9A Pending CN112245454A (zh) | 2020-10-16 | 2020-10-16 | 一种细胞疤痕修复方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112245454A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114712398A (zh) * | 2022-04-18 | 2022-07-08 | 海口仁术皮肤科门诊部有限公司 | 一种治疗凹陷性疤痕毛囊干细胞组织泥的制备方法 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU6638898A (en) * | 1997-03-05 | 1998-11-13 | Gho Holding B.V. | Method for the propagation of hair |
US20080138324A1 (en) * | 1999-11-05 | 2008-06-12 | Kleinsek Donald A | Hair mesenchymal cells |
CN108611321A (zh) * | 2018-05-11 | 2018-10-02 | 深圳市旷逸生物科技有限公司 | 一种促进皮肤创伤愈合的多因子载体的制备及应用 |
CN110475563A (zh) * | 2018-03-05 | 2019-11-19 | 杨芷 | 一种用于抗衰老修复的干细胞制剂及其制备方法 |
CN111202750A (zh) * | 2020-03-06 | 2020-05-29 | 刘景卫 | 毛囊干细胞移植术治疗疤痕过程中离体毛囊的制备方法 |
-
2020
- 2020-10-16 CN CN202011110667.9A patent/CN112245454A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU6638898A (en) * | 1997-03-05 | 1998-11-13 | Gho Holding B.V. | Method for the propagation of hair |
US20080138324A1 (en) * | 1999-11-05 | 2008-06-12 | Kleinsek Donald A | Hair mesenchymal cells |
CN110475563A (zh) * | 2018-03-05 | 2019-11-19 | 杨芷 | 一种用于抗衰老修复的干细胞制剂及其制备方法 |
CN108611321A (zh) * | 2018-05-11 | 2018-10-02 | 深圳市旷逸生物科技有限公司 | 一种促进皮肤创伤愈合的多因子载体的制备及应用 |
CN111202750A (zh) * | 2020-03-06 | 2020-05-29 | 刘景卫 | 毛囊干细胞移植术治疗疤痕过程中离体毛囊的制备方法 |
Non-Patent Citations (1)
Title |
---|
张迪等: "胎盘间充质干细胞研究进展", 《安徽医学》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114712398A (zh) * | 2022-04-18 | 2022-07-08 | 海口仁术皮肤科门诊部有限公司 | 一种治疗凹陷性疤痕毛囊干细胞组织泥的制备方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN106318904B (zh) | 间充质干细胞条件培养基用于美容品领域的用途 | |
CN110037979B (zh) | 含羊间充质干细胞外泌体冻干粉及其制备方法 | |
CN110269833A (zh) | 一种脐带间充质干细胞制剂及其制备方法和应用 | |
CN107126556B (zh) | 一种干细胞提取物及其制备方法与在制备皮肤创面修复制剂中的应用 | |
CN111202749A (zh) | 一种具有肌肉细胞修复功能的干细胞活性因子组合物的制备方法 | |
CN112891297A (zh) | 一种改善皮肤光泽度的蛋白复配液及制备方法 | |
CN115627256B (zh) | 毛囊细胞构成的多层组织工程皮肤及其制备方法与应用 | |
CN113943699B (zh) | 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 | |
WO2024120208A1 (zh) | 一种促进毛发生长的脐带间充质干细胞定向分泌组的制备方法和应用 | |
CN113041206A (zh) | 一种含间充质干细胞因子的精华液及其制备方法 | |
CN112245454A (zh) | 一种细胞疤痕修复方法 | |
CN116831983B (zh) | 一种浓缩生长因子复合凝胶制剂、制备方法及生发应用 | |
CN107779430A (zh) | 脐带间充质干细胞上清液的收集方法 | |
WO2018150440A1 (en) | Stem cell conditioned media for clinical and cosmetic applications | |
Shafiee et al. | Stem cell transplantation therapy for diabetic foot ulcer: a narrative review | |
CN110840818A (zh) | 一种美容抗衰制剂及其制备方法 | |
CN113880961B (zh) | 葡甘六糖、其制备方法、其应用和毛发再生制剂 | |
CN115025035A (zh) | 一种冻干粉、制备方法和面膜 | |
CN113621563A (zh) | 一种成纤维细胞外泌体及其制备方法与应用 | |
CN111110832A (zh) | 一种蜗牛提取液和枸骨提取液治愈创伤及疤痕的制剂及其制备方法 | |
CN112773803A (zh) | 一种小分子在制备促进皮肤伤口愈合的药物中的应用 | |
EP2040754B1 (fr) | Utilisation de fractions cellulaires du tissu adipeux pour la regeneration tissulaire post irradiation | |
CN107441481B (zh) | 一种治疗单纯皮肤损伤的经血干细胞制剂及其制备方法 | |
CN113181219A (zh) | 用于除皱的细胞制剂及其制备方法 | |
CN110755453A (zh) | 一种用于创口皮肤修复的富含hEGF的冻干粉的制备方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210122 |