CN107802891A - Organization engineering skin and preparation method thereof - Google Patents
Organization engineering skin and preparation method thereof Download PDFInfo
- Publication number
- CN107802891A CN107802891A CN201711097746.9A CN201711097746A CN107802891A CN 107802891 A CN107802891 A CN 107802891A CN 201711097746 A CN201711097746 A CN 201711097746A CN 107802891 A CN107802891 A CN 107802891A
- Authority
- CN
- China
- Prior art keywords
- cell
- skin
- support
- organization engineering
- corium
- 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
- 230000008520 organization Effects 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title abstract description 11
- 210000004027 cell Anatomy 0.000 claims abstract description 89
- 210000000130 stem cell Anatomy 0.000 claims abstract description 36
- 210000002514 epidermal stem cell Anatomy 0.000 claims abstract description 34
- 239000000203 mixture Substances 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 23
- 230000002500 effect on skin Effects 0.000 claims description 20
- 239000006285 cell suspension Substances 0.000 claims description 16
- 150000004676 glycans Chemical class 0.000 claims description 14
- 229920001282 polysaccharide Polymers 0.000 claims description 14
- 239000005017 polysaccharide Substances 0.000 claims description 14
- 238000000926 separation method Methods 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 9
- 239000011159 matrix material Substances 0.000 claims description 7
- 102000008186 Collagen Human genes 0.000 claims description 4
- 108010035532 Collagen Proteins 0.000 claims description 4
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 4
- 229920001436 collagen Polymers 0.000 claims description 4
- -1 polyoxyethylene Polymers 0.000 claims description 4
- 238000004114 suspension culture Methods 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 2
- 239000008055 phosphate buffer solution Substances 0.000 claims description 2
- 229920000954 Polyglycolide Polymers 0.000 claims 2
- 229920000747 poly(lactic acid) Polymers 0.000 claims 2
- 239000004633 polyglycolic acid Substances 0.000 claims 2
- 239000004626 polylactic acid Substances 0.000 claims 2
- 210000003491 skin Anatomy 0.000 abstract description 118
- 210000003780 hair follicle Anatomy 0.000 abstract description 15
- 230000000694 effects Effects 0.000 abstract description 11
- 210000001732 sebaceous gland Anatomy 0.000 abstract description 11
- 231100000241 scar Toxicity 0.000 abstract description 8
- 230000015572 biosynthetic process Effects 0.000 abstract description 5
- 208000028990 Skin injury Diseases 0.000 abstract description 4
- 230000004215 skin function Effects 0.000 abstract description 4
- 230000036548 skin texture Effects 0.000 abstract description 4
- 230000037314 wound repair Effects 0.000 abstract description 2
- 210000001519 tissue Anatomy 0.000 description 23
- 208000027418 Wounds and injury Diseases 0.000 description 20
- 206010052428 Wound Diseases 0.000 description 19
- 101150038679 skp gene Proteins 0.000 description 19
- 210000001339 epidermal cell Anatomy 0.000 description 17
- 210000000056 organ Anatomy 0.000 description 13
- 210000002615 epidermis Anatomy 0.000 description 11
- 210000002950 fibroblast Anatomy 0.000 description 10
- 210000004209 hair Anatomy 0.000 description 9
- 239000000463 material Substances 0.000 description 7
- 230000008929 regeneration Effects 0.000 description 6
- 238000011069 regeneration method Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 230000003321 amplification Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 230000003661 hair follicle regeneration Effects 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 238000003199 nucleic acid amplification method Methods 0.000 description 5
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- YRQNKMKHABXEJZ-UVQQGXFZSA-N chembl176323 Chemical compound C1C[C@]2(C)[C@@]3(C)CC(N=C4C[C@]5(C)CCC6[C@]7(C)CC[C@@H]([C@]7(CC[C@]6(C)[C@@]5(C)CC4=N4)C)CCCCCCCC)=C4C[C@]3(C)CCC2[C@]2(C)CC[C@H](CCCCCCCC)[C@]21C YRQNKMKHABXEJZ-UVQQGXFZSA-N 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 108010007093 dispase Proteins 0.000 description 4
- 238000001976 enzyme digestion Methods 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 210000004927 skin cell Anatomy 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 210000004207 dermis Anatomy 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 238000010231 histologic analysis Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 2
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 210000005175 epidermal keratinocyte Anatomy 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 230000037311 normal skin Effects 0.000 description 2
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 230000008733 trauma Effects 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 206010014418 Electrolyte imbalance Diseases 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003317 double-positive, alpha-beta immature T lymphocyte Anatomy 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 210000003953 foreskin Anatomy 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000000442 hair follicle cell Anatomy 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 150000003180 prostaglandins Chemical class 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 210000002374 sebum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 210000002107 sheath cell Anatomy 0.000 description 1
- 206010040882 skin lesion Diseases 0.000 description 1
- 231100000444 skin lesion Toxicity 0.000 description 1
- 230000037204 skin physiology Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000000106 sweat gland Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3886—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells comprising two or more cell types
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3895—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Dermatology (AREA)
- Transplantation (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Zoology (AREA)
- Botany (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Developmental Biology & Embryology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention discloses organization engineering skin and preparation method thereof, wherein the organization engineering skin is made up of seed cell and support, the seed cell is planted in the rack surface, and the seed cell is the mixture of corium stem cell and epidermal stem cells.The renewable hair follicle of organization engineering skin and sebaceous glands of the present invention, relative to existing organization engineering skin, closer to natural skin, the effect regenerated for skin injury wound repair, skin texture is more preferable, and the formation of scene scar can be substantially reduced, effectively recover skin function.
Description
Technical field
The present invention relates to human tissue engineering technical field, and in particular to organization engineering skin preparing technical field, especially
It is to be related to a kind of organization engineering skin of renewable hair follicle and sebaceous glands and preparation method thereof.
Background technology
Skin is first of barrier that human body resists environmental damage, and skin is the organ that human body is maximum, most complicated, and is burnt
Hinder most easily damaged organ during wound, large skin defect can cause fluid loss, Electrolyte imbalance and low albumen
Mass formed by blood stasis, severe infections etc., and dermatoplasty is the key for solving this problem, but due to autologous and heterogenous skin source and application
It is restricted in some cases, people always search for preferable Graftskin.Skin tissue engineering is to be hopeful to solve
One of approach of this problem.Skin tissue engineering is an emerging frontier branch of science, is at present closest to successful histology
Product.It is using biology and engineering principles research, is constructed for repairing, maintaining and improving injury tissue function
Tissue substituent --- organization engineering skin, its core are to create a kind of three dimensional growth support, and seed cell is (main at present
It is epidermal cell or fibroblast) external compound criteria is carried out, the Dermis equivalent or skin equivalent of artificial regeneration are formed,
Transplant at the skin lesion for needing to repair, rebuilding.
However, current organization engineering skin still has much room for improvement.
The content of the invention
It is contemplated that at least solves one of technical problem present in prior art.Therefore, one object of the present invention
It is to propose a kind of new organization engineering skin.
It should be noted that the present invention is following discovery and work based on inventor and completed：
Organization engineering skin is made up of seed cell and support.Seed cell, timbering material and cell and timbering material
The mode of interaction is to build 3 fundamentals of organization engineering skin.And the research of seed cell is always organizational project
One of focus of research, preferable seed cell should have the characteristics that：1. there is high proliferation ability and a variety of differentiation potentials；②
It is easy to obtain, and materials person is damaged small；3. the seed cell obtained can be expanded largely in vitro.Planted in organizational project careful
The research of born of the same parents is to limit the bottleneck that organizational project develops rapidly, selects which kind of cell to turn into as skin seed cells and currently grinds
The key studied carefully.
Seed cell currently used for building organization engineering skin has human epidermal stem cell, dermal fibroblast etc..Table
Skin stem cell is the tissue specifc stem cells of skin, and primary epidermal ridge is focused primarily upon in foetal period, when being extremely grown up in the form of sheets
It is distributed in basal layer of epidermis.Research shows that epidermal stem cells account for basal layer of epidermis cell 1%-10%, and maintaining, skin physiology is new
Played an important role in old metabolism.As the age increases, stem cell population is reduced, and this is also that children's skin wound healing is better than adult
One of major reason of people.Found however, inventor studies, although cultured epidermal cell is applied to clinic abroad, with
Seed cell of the epidermal cell as organization engineering skin, the shortcomings that cultivation cycle is long, scar is serious after wound healing be present, no
Functional rehabilitation can be reached.Later people develop the side that Autologous epidermis adds Allogeneic acellular dermal matrite or artificial dermis combined transplantation again
Method, but because survival rate is relatively low, artificial skin's degradation speed is too fast etc., reason is still unsatisfactory at present.Dermal fibroblast is as true
Important part of the cell in skin, can secrete cytokine profiles, as HGF, keratinocyte growth because
Son, insulin-like growth factor, TGF-0 1, prostaglandin etc., growth to epidermal cell, dividing a word with a hyphen at the end of a line and breaking up has promotion to make
With.Dermal fibroblast can promote propagation and the differentiation of epidermal stem cells as seed cell, be advantageous to the healing of the surface of a wound,
But the effect for repairing skin trauma is limited.
For example, the state-of-the-art cellularised organization engineering skin containing Autologous epidermis and corium of current performance, is being transplanted to wound
After mouthful can flap coverage, form permanent Autologous epidermis layer, isolation body tissue and extraneous barrier action served, one
Determine itself alternative skin-grafting in degree.Such organization engineering skin is on spongy collagen-glutinous polysaccharide support (C-GAG), first
The autologous dermal fibroblast of the patient through amplification cultivation is inoculated with, then again in disease of the surface seeding through amplification cultivation of support
The autologous Eponychium cell (epidermal cell) of people, the organization engineering skin containing cornified epidermal layer is formed through further culture.So
And inventor has found, this organization engineering skin does not have the power of regeneration of skin accessory organ, and the skin newly formed is hairless
The skin accessory organs such as capsule, sebaceous glands, sweat gland, lack many functions of normal skin.
Thus, inventor has carried out series of theories research and experimental exploring, it is intended to finds this organization engineering skin not
The reason for skin accessory organ can be regenerated, to solve the problem.Finally, inventor successfully has found, this organizational project skin
Skin can not regenerate one of the reason for skin accessory organ be as support (induction) cell dermal fibroblast do not have
The ability for inducing the skin accessory organs such as hair follicle to be formed, in other words, the organization engineering skin lacks induces energy with accessory organ
Cell (skin accessory organ, the progenitor cells of one layer of pluripotency when occurring originating from skin molds, in some dermal cells of power
Induction under form accessory organ's structure, and extend to dermal partial).
And then inventor is further studied, it is intended to finds a kind of with skin accessory organs such as induction hair follicles
The cell replacement dermal fibroblast of Forming ability, solves the above problems.Also, inventor surprisingly has found, dermis of skin
Stem cell (skin-derived precursors, SKPs) has skin accessory organ's Forming abilities such as induction hair follicle.SKPs is
A kind of multipotential stem cell, extend the dermal sheath cell to be formed from hair follicle hair papilla cell (DP) and DP cells, can
Grown in serum free medium in suspension cell ball.After SKPs is expelled to Immune deficient mice skin, it can induce and form hair.Mesh
It is preceding also there is not yet preparing the technology of organization engineering skin using SKPs and epidermal cell as seed cell.And inventor passes through experiment
Checking and exploration discovery, renewable hair follicle and sebum can be effectively prepared using SKPs and epidermal cell as seed cell
The organization engineering skin of gland.
Thus, in the first aspect of the present invention, the invention provides a kind of organization engineering skin, the organization engineering skin
It is made up of seed cell and support, the seed cell is planted in the rack surface.According to an embodiment of the invention, the kind
Daughter cell is the mixture of corium stem cell and epidermal stem cells.It is surprisingly found by the inventors that organization engineering skin of the invention
Renewable hair follicle and sebaceous glands, relative to existing organization engineering skin, closer to natural skin, repaiied for the skin injury surface of a wound
Multiple, skin texture regeneration effect is more preferable, and can substantially reduce the formation of scene scar, effectively recovers skin function.
, wherein it is desired to explanation, epidermal stem cells, which have, is differentiated to form epidermal cell, Hair follicle epithelial cells and sebaceous glands
The ability of epithelial cell.Thus, using the mixture of corium stem cell and epidermal stem cells as seed cell, the tissue work of acquisition
Journey skin is closer to natural skin, for can effectively recover the function of skin after dermatoplasty.
According to an embodiment of the invention, the corium stem cell and the epidermal stem cells compare 0.5-5 according to cell quantity:
1 ratio mixing.
According to an embodiment of the invention, the corium stem cell and the epidermal stem cells derive from autologous or heterogenous skin
Tissue.According to some specific examples of the present invention, the corium stem cell and the epidermal stem cells can derive from autologous skin
Skin tissue, and may be derived from any position；It can also be separated and obtained with heterogenous skin tissue, still, the tissue work finally obtained
Journey skin can only be transplanted to immunocompromised subject, or the individual immune rejection for receiving to transplant is suppressed.In addition, suitable for one kind
The organization engineering skin of animal skin transplanting, the animal varieties is not limited in the source of its seed cell, such as application on human skin transplanting can
To be used as seed cell using the corium stem cell and epidermal stem cells of other animals.
According to an embodiment of the invention, the corium stem cell and the epidermal cell are P0-P5 for cell.
According to an embodiment of the invention, the support is selected from spongy collagen-glutinous polysaccharide support, PLA/PVOH
Any one of sour support, polyoxyethylene support and acellular dermal matrix.Wherein, acellular dermal matrix (ADM) is allosome
Or xenogeneic skin passes through chemical means and carries out the dermal matrix that de- cell is prepared.ADM has complete basement membrane structure, can
To promote the propagation of epidermal cell, fibroblast and endothelial cell and differentiation.Also, the tissue compatible of acellular dermal matrix
Property is preferable, is approached with normal skin, and immunological rejection is smaller during for dermatoplasty.And spongy collagen-glutinous polysaccharide support,
Histocompatbility is preferable, and degradation speed is suitable, and preferable timbering material.
According to an embodiment of the invention, the planting density of the seed cell is 100,000-20,000,000 cell/flat
Square centimetre.Thus, the holding suitable for cell propagation and hair follicle related gene.
According to an embodiment of the invention, by the way that the cell suspension of seed cell is added dropwise into the side on support and being incubated
Formula realizes the plantation.Thus, planting effect is good, and the organization engineering skin structure of acquisition is close to natural skin.
In the second aspect of the present invention, present invention also offers a kind of method for preparing organization engineering skin.According to this hair
Bright embodiment, this method is using the mixture of corium stem cell and epidermal cell as seed cell, and by the seed cell
Plant on support, to obtain the organization engineering skin.Organized it should be noted that the method for the present invention is different from tradition
Engineering skin preparation method.The preparation method of traditional organizational project is that dermal cell and epidermal cell is suitable in different time layering
Sequence is planted, and generally first plants dermal cell, plants cuticular cellulose above it again after cultivating a period of time, and carry out one again
The culture of section time, it needs two kinds of different cultivating systems, and method is complicated, and technical requirements are high.And the present invention method be by
Epidermal stem cells and the disposable plantation of corium stem cell mixing, it is easy to operate, and planting effect is good.Also, inventor is in surprise
It was found that epidermal stem cells and the plantation of corium stem cell form skin knot to position needed for after timbering material, oneself can reaching
Structure, hair follicle and sebaceous glands are regenerated, it, closer to natural skin, is created relative to existing organization engineering skin for skin injury
Face is repaired, the effect of skin texture regeneration is more preferable, and can substantially reduce the formation of scene scar, effectively recovers skin function.
In addition it is also necessary to, it is emphasized that the method for the present invention, the organization engineering skin prepared have appendicle (hair
Capsule, sebaceous glands) power of regeneration.Existing patented technology needs to be inoculated with (dry) cell of these organs respectively, is such as isolated from hair follicle
Cell, be isolated from the cell of sebaceous glands.However, it is extremely difficult to separate these cells, and cultured and amplified in vitro is difficult to, because
And such technology does not have practicality actually.And the present invention use seed cell --- corium stem cell and epidermal stem are thin
Born of the same parents, separation easily and can amplification in vitro, be easy to put into practice and promote, practical value is high.
According to an embodiment of the invention, the corium stem cell and the epidermal stem cells compare 0.5-5 according to cell quantity:
1 ratio mixing.
According to an embodiment of the invention, the corium stem cell and the epidermal stem cells derive from autologous or heterogenous skin
Tissue.
According to an embodiment of the invention, the corium stem cell and the epidermal cell are P0-P5 for cell.Thus,
The organization engineering skin activity and better function of acquisition.
According to an embodiment of the invention, the support is selected from spongy collagen-glutinous polysaccharide support, PLA/PVOH
Any one of sour support, polyoxyethylene support and acellular dermal matrix.Thus, the organization engineering skin of acquisition is used to transplant
When histocompatbility it is good, degradation speed matters.
According to an embodiment of the invention, the planting density of the seed cell is 100,000-20,000,000 cell/flat
Square centimetre.As it was previously stated, this density is suitable to the holding of cell propagation and hair follicle related gene.
According to an embodiment of the invention, by the way that the cell suspension of seed cell is added dropwise into the side on support and being incubated
Formula realizes the plantation.Thus, planting effect is good, and the organization engineering skin structure of acquisition is close to natural skin.
According to an embodiment of the invention, the cell suspension is hanged using corium stem cell media or phosphate buffer solution
What the floating seed cell obtained, and the concentration of the cell suspension is 2.0 × 104-2.0×109Cells/ml.
According to an embodiment of the invention, in 37 DEG C, 5%CO2Under conditions of, carry out described be incubated -14 days 4 hours.Thus,
Incubation effect is good, the organization engineering skin activity and better function of acquisition, closer to natural skin.
According to other embodiments of the present invention, the described method comprises the following steps：
(1) spongy collagen-glutinous polysaccharide support is prepared using the acetum of glutinous polysaccharide and the acetum of collagen；
(2) corium stem cell is obtained from skin histology separation, and carries out suspension culture；
(3) epidermal stem cells are obtained from skin histology separation, and carries out adhere-wall culture；
(4) will through suspension the corium stem cell cultivated and the epidermal stem cells Jing Guo adhere-wall culture, according to cell quantity
Compare 0.5-5:1 ratio mixing, obtains seed cell existing in the form of cell suspension, then according to 100,000-20,000,
The planting density of 000 cells/square cm, the cell suspension is added dropwise on the spongy collagen-glutinous polysaccharide support, and
In 37 DEG C, 5%CO2Under conditions of cultivate -14 days 4 hours, so as to which seed cell is planted on support, obtaining the tissue work
Journey skin.
Thus, the renewable hair follicle of organization engineering skin and sebaceous glands prepared, histocompatbility is good, relative to existing
Organization engineering skin, closer to natural skin, the effect regenerated for skin injury wound repair, skin texture is more preferable, and
The formation of scene scar can be substantially reduced, effectively recovers skin function.
According to some specific examples of the present invention, the method for preparing organization engineering skin of the invention can also include following
Step：
(1) C-GAG preparation：
It is 1.0- that the acetum for the glutinous polysaccharide that 2mL mass fractions are 1.0-2.0% is added drop-wise into 25mL mass fractions
In the acetum of 2.0% collagen, 15000rpm stirs 4h at 4 DEG C, and 3500rpm centrifuges 5min, molten to obtain C-GAG
Liquid；C-GAG solution is poured into homemade mould, then mould is put into -10 DEG C to -80 DEG C of refrigerator and (uses isopropanol
Control cooling rate be 1 DEG C/min for bath), after 12h, taking-up mould, 24h is dried in vacuo in freeze dryer, then will be dried
C-GAG branch, which is placed in vacuum drying chamber, is heat-treated 24h, and heat treatment temperature is 120 DEG C, vacuum 40Pa；
(2) SKPs separation and culture：
Skin tissue sample is taken, with the 4 DEG C of digestion of 0.1%-1%dispase II enzymes overnight, separates epidermis and skin corium,
With 0.1%-1%Collagenase I enzymic digestion dermal tissues, through 40 μm of aperture cell sieve filterings, 1300rpm centrifugations 5min is received
Collect cell (SKPs), then with containing B27 and adding EGF and bFGF DMEM/F12 (3:1) culture medium, in 37 DEG C of 5%CO2Training
Support in case and suspension culture is carried out to SKPs；
(3) separation and culture of epidermal stem cells：
Skin tissue sample is taken, with the 4 DEG C of digestion of 0.1%-1%dispase II enzymes overnight, separates epidermis and skin corium；
With 0.1%-1%Collagenase I enzymic digestions epidermal tissue, through 40 μm of aperture cell sieve filterings, 1300rpm centrifugations 5min is received
Collect cell, with CNT-07 epidermal keratinocyte culture mediums, in 37 DEG C, 5%CO2Incubator in epidermal stem cells cell carry out
Adhere-wall culture；
(4) can hair follicle regeneration organization engineering skin preparation：
The epidermal stem cells in the SKPs and P0-P5 generations in P0-P5 generations are collected respectively, by SKPs and epidermal stem cells according to quantity
Compare 0.5-5:1 mixing, obtains cell suspension, cell suspension is added dropwise on C-GAG supports, planting density 100,000-20,
000,000 cells/square cm, culture -14 days 4 hours is then incubated, produces organization engineering skin.
Furthermore, it is necessary to explanation, by the organization engineering skin be used for skin trauma recover transplanting when, can according to
Operation is carried out：Organization engineering skin is migrated into wound, then covers not mucous membrane and 3M ventilated membranes successively, with self-adhering-type bandage
Bind up a wound.It is surface of a wound recovery and the hair follicle regeneration situation of observable wound after 3 weeks.
Furthermore, it is necessary to explanation, organization engineering skin of the invention and preparation method thereof have following advantages at least
One of：
1st, power of regeneration is strong：After the organization engineering skin transplanting of the present invention, epidermis is not only formed, also renewable formation hair
Capsule, sebaceous glands, obvious scar is not formed.
2nd, prepare simple：Need not be by dermal cell and (other) plantation of epidermal cell layering and culture.
3rd, cell derived is extensive and can cultivate amplification：Key cells of the present invention, corium source of human stem cell is extensive, and
Still there is the ability that the structures such as induction hair follicle are formed after culture amplification.
The additional aspect and advantage of the present invention will be set forth in part in the description, and will partly become from the following description
Obtain substantially, or recognized by the practice of the present invention.
Brief description of the drawings
The above-mentioned and/or additional aspect and advantage of the present invention will become in the description from combination accompanying drawings below to embodiment
Substantially and it is readily appreciated that, wherein：
Fig. 1 is the organization engineering skin support C-GAG prepared in the step 1 of embodiment 1 scanning electron microscope (SEM) photograph；
Fig. 2 is the control group and experimental group surface of a wound hair comparison diagram of embodiment 2；
Fig. 3 is the control group and experimental group fabric analysis comparison diagram of embodiment 2.
Embodiment
The solution of the present invention is explained below in conjunction with embodiment.It will be understood to those of skill in the art that following
Embodiment is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Unreceipted particular technique or bar in embodiment
Part, carried out according to the technology described by document in the art or condition or according to product description.Agents useful for same or instrument
The unreceipted production firm person of device, being can be by the conventional products of acquisition purchased in market.
Embodiment 1：Prepare can hair follicle regeneration organization engineering skin
With reference to the present invention the method for preparing organization engineering skin, prepare can hair follicle regeneration organization engineering skin.Specifically
It is as follows：
(1)：C-GAG preparation
The acetum for the glutinous polysaccharide that 2mL mass fractions are 1.6% is added drop-wise to the collagen that 25mL mass fractions are 1.3%
Acetum in, at 4 DEG C 15000rpm stir 4h, 3500rpm centrifugation 5min, C-GAG solution is poured into homemade mould
In, it is put into -10 DEG C to -80 DEG C of refrigerator and (uses isopropanol bath to control cooling rate as 1 DEG C/min), after 12h, takes out mould
Tool, is dried in vacuo 24h in freeze dryer, and dried C-GAG branch is placed in vacuum drying chamber and is heat-treated 24h, temperature 120
DEG C, vacuum 40Pa, produce C-GAG supports (its scanning electron microscope (SEM) photograph is shown in Fig. 1).
(2)：SKPs separation and culture
Take C57 newborn rat Back skin samples, with the digestion of 4 DEG C of 0.3%dispase II enzymes overnight, separation epidermis with it is true
Cortex, with 0.35%Collagenase I enzymic digestion dermal tissues, through 40 μm of aperture cell sieve filterings, 1300rpm centrifuges 5min
Cell (SKPs) is collected, 20ng/mL EGF and 40ng/mL bFGF DMEM/F12 (3 are with the addition of with (1%) containing B27:1) cultivate
Base, in 37 DEG C, 5%CO2Incubator in suspension culture is carried out to SKPs；
(3)：The separation and culture of epidermal cell
Take adult's foreskin to take skin tissue sample, with the digestion of 4 DEG C of 1%dispase II enzymes overnight, separation epidermis with it is true
Cortex, with 0.35%Collagenase I enzymic digestions epidermal tissue, through 40 μm of aperture cell sieve filterings, 11300rpm is centrifuged
5min collects cell (epidermal stem cells), with CNT-07 epidermal keratinocyte culture mediums, in 37 DEG C, 5%CO2Incubator in it is right
Epidermal stem cells carry out adhere-wall culture.
(4)：Can hair follicle regeneration organization engineering skin preparation
The epidermal cell in the SKPs and P4 generations in P4 generations is collected respectively, by two kinds of cells according to SKPs:Epidermal cell=2:1 enters
Row mixing, obtains cell suspension, and cell suspension is added dropwise on C-GAG supports, and the cell of planting density 5,000,000/square
Centimetre, in 37 DEG C, 5%CO2Incubator in carry out be incubated culture 7 days, produce organization engineering skin, it is standby.
Embodiment 2
Method according to embodiment 1 prepares organization engineering skin, differs only in：
SKPs:Epidermal stem cells mixed proportion is 0.5:1, the cells/square cm of planting density 100,000, it is incubated training
It is 4 hours to support the time.
Embodiment 3
Method according to embodiment 1 prepares organization engineering skin, differs only in：
SKPs:Epidermal stem cells mixed proportion is 5:1, the cells/square cm of planting density 20,000,000, it is incubated training
It is 14 days to support the time.
Comparative example 1
Method according to embodiment 1 prepares organization engineering skin, differs only in：
SKPs, and delete step (2) are substituted with dermal fibroblast.Wherein, acquisition is separately cultured according to known technique
Dermal fibroblast.
Embodiment 4：The foundation of the full thick-layer wound transplantation model of BALB/c nu/nu mouse
The organization engineering skin obtained for embodiment 1-3 and comparative example 1, follows the steps below transplanting respectively：
By BALB/c nu/nu mouse anesthesias, the full thick-layer wound of 1 10mm diameter is made at its back, by the tissue
Engineering skin migrates to wound, then covers the not (article No.s of mucous membrane Smith＆NephewConformant 2 successively：5955044)
With 3M ventilated membrane Tegaderm (article No.s：1624w), bound up a wound with self-adhering-type bandage.After 3 weeks, mouse is put to death, observes hair
Quantity and progress histologic analysis.
That is, experiment sets 3 experimental groups and 1 control group, the organization engineering skin that wherein control group uses is pair
What ratio 1 prepared (relative to the organization engineering skin of embodiment 1, differs only in SKPs and replaces in order to which corium is into fiber
Cell).
As a result find, the organization engineering skin of embodiment 1-3 and comparative example 1 is used for after transplanting, whole wounds without infection,
Wound, which heals, to be finished.Surface of a wound recovery situation and hair formational situation no significant difference between experimental group, and the wound of control group
Face recovery situation and hair formational situation are substantially poor.
Specifically：
1) control group：The surface of a wound formed scar, it is hairless go out (see Fig. 2), histologic analysis show cambium without skin
Accessory structure is generated (see Fig. 3).
2) experimental group (by taking the experimental group for the organization engineering skin that embodiment 1 prepares as an example)：The surface of a wound without obvious scar,
And visible a large amount of black hairs are born (see Fig. 2), it is attached that histologic analysis shows that cambium forms hair follicle and sebaceous glands etc.
Structure (see Fig. 3).
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or the spy for combining the embodiment or example description
Point is contained at least one embodiment or example of the present invention.In this manual, to the schematic representation of above-mentioned term not
Necessarily refer to identical embodiment or example.Moreover, specific features, structure, material or the feature of description can be any
One or more embodiments or example in combine in an appropriate manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that：Not
In the case of departing from the principle and objective of the present invention a variety of change, modification, replacement and modification can be carried out to these embodiments, this
The scope of invention is limited by claim and its equivalent.
Claims (10)
1. a kind of organization engineering skin, is made up of seed cell and support, the seed cell is planted in the rack surface, its
It is characterised by, the seed cell is the mixture of corium stem cell and epidermal stem cells.
2. organization engineering skin according to claim 1, it is characterised in that the corium stem cell and the epidermal stem are thin
Born of the same parents compare 0.5-5 according to cell quantity:1 ratio mixing,
Optionally, the corium stem cell and the epidermal stem cells derive from autologous or heterogenous skin tissue,
Optionally, the corium stem cell and the epidermal stem cells are P0-P5 for cell,
Optionally, the support is selected from spongy collagen-glutinous polysaccharide support, polylactic acid/polyglycolic acid support, polyoxyethylene branch
Any one of frame and acellular dermal matrix.
3. organization engineering skin according to claim 1, it is characterised in that the planting density of the seed cell is 100,
000-20,000,000 cells/square cm,
Optionally, the plantation is realized on support and by way of being incubated by the way that the cell suspension of seed cell is added dropwise.
A kind of 4. method for preparing organization engineering skin, it is characterised in that with corium stem cell and the mixture of epidermal stem cells
Planted as seed cell, and by the seed cell on support, to obtain the organization engineering skin.
5. according to the method for claim 4, it is characterised in that the corium stem cell and the epidermal stem cells are according to thin
Born of the same parents' quantity compares 0.5-5:1 ratio mixing,
Optionally, the corium stem cell and the epidermal stem cells derive from autologous or heterogenous skin tissue,
Optionally, the corium stem cell and the epidermal stem cells are P0-P5 for cell,
Optionally, the support is selected from spongy collagen-glutinous polysaccharide support, polylactic acid/polyglycolic acid support, polyoxyethylene branch
Any one of frame and acellular dermal matrix.
6. according to the method for claim 4, it is characterised in that the planting density of the seed cell is 100,000-20,
000,000 cells/square cm.
7. according to the method for claim 4, it is characterised in that by the way that the cell suspension of seed cell is added dropwise on support
And the mode being incubated realizes the plantation.
8. according to the method for claim 7, it is characterised in that the cell suspension be using corium stem cell media or
Phosphate buffer solution suspends what the seed cell obtained, and the concentration of the cell suspension is 2.0 × 104-2.0×109Carefully
Born of the same parents/milliliter.
9. according to the method for claim 7, it is characterised in that in 37 DEG C, 5%CO2Under conditions of, it is small to carry out the incubation 4
When -14 days.
10. according to the method for claim 4, it is characterised in that the described method comprises the following steps：
(1) spongy collagen-glutinous polysaccharide support is prepared using the acetum of glutinous polysaccharide and the acetum of collagen；
(2) corium stem cell is obtained from skin histology separation, and carries out suspension culture；
(3) epidermal stem cells are obtained from skin histology separation, and carries out adhere-wall culture；
(4) will through suspension the corium stem cell cultivated and the epidermal stem cells Jing Guo adhere-wall culture, according to 0.5-5:1 ratio
Mixing, obtains seed cell existing in the form of cell suspension, then according to 100,000-20,000,000 cells/square cms
Planting density, the cell suspension is added dropwise on the spongy collagen-glutinous polysaccharide support, and in 37 DEG C, 5%CO2's
Under the conditions of cultivate -14 days 4 hours, so as to which seed cell is planted on support, obtaining the organization engineering skin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711097746.9A CN107802891A (en) | 2017-11-09 | 2017-11-09 | Organization engineering skin and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711097746.9A CN107802891A (en) | 2017-11-09 | 2017-11-09 | Organization engineering skin and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107802891A true CN107802891A (en) | 2018-03-16 |
Family
ID=61583106
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711097746.9A Pending CN107802891A (en) | 2017-11-09 | 2017-11-09 | Organization engineering skin and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107802891A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110804580A (en) * | 2019-11-21 | 2020-02-18 | 山东大学 | Culture method for forming early micro hair follicle in vitro and application thereof |
| CN110862954A (en) * | 2018-08-28 | 2020-03-06 | 广州洁特生物过滤股份有限公司 | Cell culture method based on three-dimensional cell culture support |
| CN112206357A (en) * | 2020-09-25 | 2021-01-12 | 清华大学 | Tissue engineering skin biological ink, preparation method, regeneration method and system |
| CN113201481A (en) * | 2021-04-19 | 2021-08-03 | 清华大学深圳国际研究生院 | Skin microsphere and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062429A (en) * | 2006-04-24 | 2007-10-31 | 中国人民解放军军事医学科学院野战输血研究所 | Method for constructing tissue engineering double-layered skin and the application thereof |
| CN101063109A (en) * | 2006-04-24 | 2007-10-31 | 中国人民解放军军事医学科学院野战输血研究所 | Construction method and application for complexion adjustable organization engineering skin |
| CN101775366A (en) * | 2010-02-05 | 2010-07-14 | 中国人民解放军第四军医大学 | Preparation method of tissue engineering skin containing hair follicles |
| CN103785064A (en) * | 2013-08-26 | 2014-05-14 | 济南磐升生物技术有限公司 | Method for regenerating human intact skin tissue by use of in vitro cultured cells |
| CN104491931A (en) * | 2014-12-02 | 2015-04-08 | 清华大学深圳研究生院 | Sebaceous gland-containing skin tissue as well as formation method and application thereof |
| CN105013019A (en) * | 2015-08-18 | 2015-11-04 | 济南磐升生物技术有限公司 | Tissue engineering full-layer skin and preparing method thereof |
-
2017
- 2017-11-09 CN CN201711097746.9A patent/CN107802891A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062429A (en) * | 2006-04-24 | 2007-10-31 | 中国人民解放军军事医学科学院野战输血研究所 | Method for constructing tissue engineering double-layered skin and the application thereof |
| CN101063109A (en) * | 2006-04-24 | 2007-10-31 | 中国人民解放军军事医学科学院野战输血研究所 | Construction method and application for complexion adjustable organization engineering skin |
| CN101775366A (en) * | 2010-02-05 | 2010-07-14 | 中国人民解放军第四军医大学 | Preparation method of tissue engineering skin containing hair follicles |
| CN103785064A (en) * | 2013-08-26 | 2014-05-14 | 济南磐升生物技术有限公司 | Method for regenerating human intact skin tissue by use of in vitro cultured cells |
| CN104491931A (en) * | 2014-12-02 | 2015-04-08 | 清华大学深圳研究生院 | Sebaceous gland-containing skin tissue as well as formation method and application thereof |
| CN105013019A (en) * | 2015-08-18 | 2015-11-04 | 济南磐升生物技术有限公司 | Tissue engineering full-layer skin and preparing method thereof |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110862954A (en) * | 2018-08-28 | 2020-03-06 | 广州洁特生物过滤股份有限公司 | Cell culture method based on three-dimensional cell culture support |
| CN110804580A (en) * | 2019-11-21 | 2020-02-18 | 山东大学 | Culture method for forming early micro hair follicle in vitro and application thereof |
| CN110804580B (en) * | 2019-11-21 | 2021-09-28 | 山东大学 | Culture method for forming early micro hair follicle in vitro and application thereof |
| CN112206357A (en) * | 2020-09-25 | 2021-01-12 | 清华大学 | Tissue engineering skin biological ink, preparation method, regeneration method and system |
| CN112206357B (en) * | 2020-09-25 | 2021-12-28 | 清华大学 | Tissue engineering skin regeneration system |
| CN113201481A (en) * | 2021-04-19 | 2021-08-03 | 清华大学深圳国际研究生院 | Skin microsphere and preparation method and application thereof |
| CN113201481B (en) * | 2021-04-19 | 2023-10-13 | 清华大学深圳国际研究生院 | Skin microsphere and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108525021B (en) | Tissue engineering skin containing blood vessels and hair follicle structures based on 3D printing and preparation method thereof | |
| CN107802891A (en) | Organization engineering skin and preparation method thereof | |
| CN104726406B (en) | It is a kind of to induce the method that dental pulp Derived from Mesenchymal Stem Cells is nerve cell | |
| CN104263699A (en) | Culture method for large-scale preparation of clinical treatment level dermal multipotent stem cells for cell transplantation | |
| CN104312970A (en) | Preparation method of clinical treatment level epidermal stem cell for cell therapy by applying human extracellular matrix screening and mass culture | |
| CN104087551B (en) | The method of a kind of Isolation and culture people's epidermal cell | |
| CN105820998A (en) | Isolation extraction and culture method for human adipose-derived stem cells (ADSCs) for clinical back-transfusion grade cell therapy | |
| CN102311942A (en) | Vitamin C induced mesenchymal stem cell membrane and preparation method thereof | |
| CN104946585A (en) | Production and freeze-drying method and application for supernate of culture solution of mesenchymal stem cell | |
| CN102367434B (en) | Amniotic membrane microcarrier capable of simulating niche microenvironment for growth of epidermal stem cells and skin substitute thereof | |
| CN107349475B (en) | The artificial organ engineering skin and preparation method thereof that nano fibrous membrane is layering with stem cell | |
| CN103275922B (en) | A kind of fine-wool sheep hair follicle stem cells isolated culture method | |
| CN104263698B (en) | Clinical treatment level cell therapy is prepared human cell's epimatrix screening and culturing method with fibroblast scale | |
| CN106620855B (en) | A kind of construction method of composite tissue engineering skin | |
| CN105255822A (en) | Method for screening and culturing extracellular hair follicle stem cell matrix for clinic treatment level cell therapy | |
| CN105079783A (en) | Pharmaceutical composition and preparation method and application thereof | |
| CN105018417B (en) | The load intrinsic stem cell of amnion freezes active amnia particle and its conditioned medium and application | |
| CN101711890A (en) | Extracellular matrix gel model used for researching development and differentiation of embryonic stem cells | |
| CN104491931B (en) | Sebaceous gland-containing skin tissue as well as formation method and application thereof | |
| CN107254431A (en) | A kind of novel tissue engineering skin preparation method | |
| CN101264344B (en) | Preparation of artificial skin containing hair follicle and artificial skin prepared by the same | |
| US20200149005A1 (en) | The method of autologous primary hair follicles preparation in 3d culture | |
| CN105238739A (en) | Selective culture method for large-scale preparation of human extracellular matrix through melanocytes for clinic treatment level cell therapy | |
| CN101063109A (en) | Construction method and application for complexion adjustable organization engineering skin | |
| CN105963795A (en) | Method for preparing tissue engineering epidermis based on collagen |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180316 |