WO2016086527A1 - Tissu cutané contenant une glande sébacée, procédé de formation et utilisation de celui-ci - Google Patents

Tissu cutané contenant une glande sébacée, procédé de formation et utilisation de celui-ci Download PDF

Info

Publication number
WO2016086527A1
WO2016086527A1 PCT/CN2015/071777 CN2015071777W WO2016086527A1 WO 2016086527 A1 WO2016086527 A1 WO 2016086527A1 CN 2015071777 W CN2015071777 W CN 2015071777W WO 2016086527 A1 WO2016086527 A1 WO 2016086527A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
stem cells
skin
dermal
final concentration
Prior art date
Application number
PCT/CN2015/071777
Other languages
English (en)
Chinese (zh)
Inventor
吴耀炯
王潇潇
王旭升
Original Assignee
清华大学深圳研究生院
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 清华大学深圳研究生院 filed Critical 清华大学深圳研究生院
Publication of WO2016086527A1 publication Critical patent/WO2016086527A1/fr

Links

Images

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
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • 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
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • 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

Definitions

  • the present invention relates to the field of cell biology and regenerative medicine, and in particular to a method of forming a sebaceous gland and its use, and more particularly to a method of forming a sebaceous gland, a skin tissue containing a sebaceous gland, and a method of forming a sebaceous gland for use in skin regeneration.
  • the skin As the largest tissue of the human body, the skin is a barrier to contact with the external environment. When external skin damage or diseases cause skin defects, skin damage and infection are often caused. China has a large population, with 15 million burns and ulcers per year, of which 3.5 million are required for skin grafts and more than 400 million square centimeters. Therefore, finding an ideal skin substitute has always been a clinically urgent problem. With the rapid development of cell biology, materials science, biochemistry, bioengineering, and transplantation, people have a deeper understanding of the biological characteristics of epidermal keratinocytes, the processing and synthesis of artificial materials, and so on. It is possible to build tissue engineering skin.
  • the sebaceous gland is the most important accessory organ for maintaining the stability of skin structure. Sebaceous glands and sweat and exfoliated epithelial cells form a layer of sebum sweat latex film on the surface of the skin. Its thickness is 7-10 ⁇ m, which is a three-dimensional lipid structure on the skin surface. Promotes the integrity of the skin barrier. In the absence of this three-dimensional cortical structure, rough skin and dry hair will appear. Since some skins of the palms, ankles, fingers, and toes do not have sebaceous glands, skin dryness often occurs.
  • tissue engineering skin has a dermis and epidermis similar to normal skin, it lacks sebaceous glands and cannot provide nutrient and moisturizing, antibacterial and anti-oxidant protection and moisture retention. After skin transplantation, skin shrinkage and deformation are serious and appearance is easy. Problems such as chapped and susceptible to infection, while the performance of tissue-engineered skin is significantly lower than that of natural normal skin.
  • the present invention aims to solve at least one of the technical problems existing in the prior art. To this end, it is an object of the present invention to provide a method of forming a sebaceous gland.
  • epidermal stem cells derived from the epidermis of the skin and dermal cells derived from the dermis of the skin have a large potential for expansion, and after isolation and culture in vitro, epidermal stem cells are characterized by high expression of integrin CD29, CD49f and K15 ( Keratin 15), and the characteristic marker molecule expression of two stem cells in subsequent passages stable.
  • the inventors amplified the above two cells in different culture systems, and then transplanted the two cells to the skin of nude mice in proportion, and histological analysis revealed a newly formed sebaceous gland structure.
  • the invention provides a method of forming a sebaceous gland.
  • the method comprises co-culturing dermal cells and epidermal stem cells in a gel.
  • the inventors have surprisingly found that the method can form skin tissue with sebaceous glands.
  • the present application forms a method of skin tissue having sebaceous glands, which is simple, convenient, quick, easy to control, and easy to achieve large-scale production.
  • the method of sebaceous gland according to the above embodiment of the present invention may further have the following additional technical features:
  • the co-cultivation is carried out subcutaneously in the animal.
  • the regeneration of the skin tissue of the sebaceous glands is promoted, and the structure of the regenerated skin tissue is similar to that of normal skin.
  • the dermal cells comprise at least one of dermal stem cells and fibroblasts.
  • the dermal cells are fibroblasts
  • the gel contains insulin, dexamethasone, glitazone and XAV939 (3,5,7,8-tetrahydro-2-[ 4-(Trifluoromethyl)phenyl]-4H-thiopyrano[4,3-d]pyrimidin-4-one).
  • the chemical agent induces differentiation of epidermal stem cells into free sebaceous glands, thereby forming a sebaceous gland structure similar to normal skin.
  • the final concentration of the insulin is 50-150 ⁇ g/mL
  • the final concentration of the dexamethasone is 10 -4 -10 -6 M
  • the final concentration of the glitazone is 150-250 ⁇ M
  • the end of the XAV939 The concentration is 5-15 ⁇ M.
  • the chemical agent induces the differentiation of epidermal stem cells and fibroblasts into free sebaceous glands.
  • the final concentration of the insulin is 100 ⁇ g / mL
  • the final concentration of the dexamethasone is 10 -5 M
  • the final concentration of the glitazone is 200 ⁇ M
  • the final concentration of the XAV939 is 10 ⁇ M.
  • the chemical agent induces the differentiation of epidermal stem cells and fibroblasts into free sebaceous glands.
  • final concentration refers to the concentration of the chemical agent in the gel after adding various chemical agents to the gel containing the mixed cells.
  • the ratio of the dermal cells to the epidermal stem cells is 10:1 to 1:10.
  • the number of mixed cells is not less than 10 4 per 3 mm 2 of skin.
  • the number of mixed cells satisfies the need for regeneration of sebaceous gland skin tissue.
  • the gel is at least one selected from the group consisting of Matrigel, collagen or hydrogel.
  • the dermal cells and the epidermal stem cells are all derived from autologous, allogeneic or xenogeneic cells. It should be noted that the xenogenic-derived cells are used for the regeneration of sebaceous glands in immunodeficient animals. Thereby, the regenerated sebaceous gland skin tissue has a small rejection effect.
  • the dermal cells and the epidermal stem cells are independently cultured in a liquid medium for 0 to 30 days, wherein the dermal stem cells are suspension culture, and the cell culture medium is DMEM/F12. (3:1), 2% B27, 40 ng/ ⁇ L bFGF and 20 ng/ ⁇ L EGF; epidermal stem cells are adherent culture, and the cell culture medium is CnT-07 medium; wherein the dermal stem cells further contain liquid medium a cell active factor and an extracellular matrix molecule, wherein the cell active factor is at least one selected from the group consisting of bFGF and EGF, and the extracellular matrix molecule is at least one selected from the group consisting of hyaluronic acid and fibronectin.
  • the dermal stem cells and epidermal stem cells have high activity and small cell variation.
  • the method further comprises: identifying the oil secretion of the surface of the skin tissue containing the regenerated sebaceous glands using in situ mass spectrometry. Thereby, the secretion of oil on the surface of the skin tissue containing the regenerated sebaceous glands can be more accurately identified.
  • the present invention also provides a skin tissue containing a sebaceous gland.
  • the sebaceous glands are formed by the method described in the foregoing embodiments of the invention. The inventors have surprisingly found that the skin tissue has a structure of sebaceous glands similar to normal skin tissue.
  • the invention also provides for the use of the aforementioned method in skin regeneration.
  • the method is for regenerating a sebaceous gland or a skin tissue containing a sebaceous gland. The inventors have surprisingly found that by applying this method in skin regeneration, sebaceous glands similar to normal skin can be obtained.
  • FIG. 1 shows a photograph of the expression result of the cell surface marker molecule CD49f of epidermal stem cells according to an embodiment of the present invention
  • FIG. 2 shows a photograph of the expression result of the cell surface marker Nestin of dermal stem cells according to an embodiment of the present invention
  • Figure 3 shows a photograph of confocal microscopy of normal skin tissue sections in accordance with one embodiment of the present invention
  • FIG. 4 is a photograph showing confocal microscopic observation of a skin tissue section formed by co-transplantation of epidermal stem cells and dermal stem cells according to an embodiment of the present invention
  • Figure 5 is a photograph showing confocal microscopic observation of skin tissue sections formed by co-transplantation of epidermal stem cells and fibroblasts after induction of co-transplantation with chemical agents according to an embodiment of the present invention
  • Figure 6 is a graph showing the results of sebaceous gland secretion detected by MALDI mass spectrometry in skin tissue formed after transplantation according to one embodiment of the present invention.
  • ex vivo skin stem cells are isolated and cultured from the skin of human C57 mice or human scalp tissues in the following examples;
  • the dermal stem cell culture medium is composed of DMEM/F12 (3: 1) (purchased from Invitrogen) containing 2% B27 (purchased from Invitrogen), 40 ng/ ⁇ L bFGF (Peprotech) and 20 ng/ ⁇ L EGF (Peprotech);
  • fibroblast culture medium is DMEM containing 10% fetal bovine serum Accutase was purchased from sigma; 3M membrane was Tegaderm TM Film; suspension cell culture dish was purchased from Guangzhou Jiete Biofiltration Products Co., Ltd., and the product number was MCD-000-090.
  • Adherent cell culture dish purchased from Corning. Matrigel was purchased from BD. The experimental animals were purchased from the Guangdong Medical Laboratory Animal Center.
  • Identification of epidermal stem cells The identification of cell surface markers is as follows: Take the first, third, and fifth representative skin cells for cell climbing, and perform immunofluorescence staining of integrin CD29, CD49f and K15. The coloration of the cells. It was observed that the integrin CD29, CD49f and K15 of epidermal stem cells were highly expressed, and the specific results are shown in Fig. 1.
  • the primary cells of the dermal stem cells were passed through a 200 mesh sieve, and then centrifuged at 300 ⁇ g for 10 min, and the supernatant was discarded, and the primary cells were suspended in the dermal stem cell culture medium.
  • the dermal stem cell culture medium suspension primary cells were seeded in an non-adherent culture dish.
  • dermal stem cells to induce epidermal stem cells to form sebaceous gland structures, the specific process is as follows:
  • Example 1 The epidermal stem cells obtained in Example 1 were cultured, and the third representative skin stem cells were digested with Accutase, and a suspension of epidermal stem cells having a total cell number of 1 million was prepared in PBS.
  • Example 2 The dermal stem cells obtained in Example 2 were cultured, and a total of 1 million cell suspensions of the third generation dermal stem cells were prepared in PBS.
  • BALB/c nude mice were anesthetized with 1% sodium pentobarbital, and a wound having an area of about 7 mm 2 was made on the skin of the back with a puncher.
  • Example 1 The epidermal stem cells obtained in Example 1 were cultured, and the third representative skin stem cells were digested with Accutase, and a cell suspension of 2 million was prepared in PBS.
  • mice were anesthetized with 1% sodium pentobarbital, and a wound having an area of about 7 mm 2 was made on the skin of the back with a puncher.
  • Example 3 and Example 4 The skin tissue regions of the stem cells transplanted in the experimental group and the control group in Example 3 and Example 4 were respectively cut out, fixed in 4% paraformaldehyde overnight, immersed in PBS for 3 hours, and dehydrated in 30% sucrose for 8 hours.
  • Tissue sections were permeabilized for 10 minutes with 0.2-0.5% triton X-100 (in PBS).
  • Tissue sections were blocked with 2% BSA for 30 minutes and then washed twice with PBS.
  • the tissue dyeing sample with good sealing is placed in a 4 degree refrigerator and can be stored for more than 2 weeks.
  • the method for identifying the oil secretion ability of the sebaceous glands of the skin tissue after transplantation in Example 4 by in situ mass spectrometry is specifically as follows:
  • mice transplanted with the stem cell skin of Example 4 were anesthetized with 1% sodium pentobarbital, and the back skin was scrubbed with 100% ethanol to wash off all the oily substances on the skin surface.
  • tissue sections were pretreated by a 50% ACN aqueous solution containing 0.1% TFA to prepare a 15 g/L CHCA matrix solution.
  • the pump was used to spray 100 ⁇ l of the substrate solution evenly on the surface of the tissue at 5 ⁇ l/s.
  • the substrate solution was sprayed again and circulated twice.
  • tissue section of the sprayed substrate was placed at room temperature, and the solvent was evaporated to precipitate crystals, which were then dried in a desiccator for 2 hours.
  • the dried tissue sections were attached to a MALDI stainless steel target plate with a conductive tape and sent to a mass spectrometer for analysis.
  • Mass spectrometry conditions Randomly take 3 points of each tissue section, add 1000 scan data to each point to obtain the mass spectrum, and add 3 points to obtain the total mass spectrum. Detection by positive ion reflection mode, mass scanning range m/z 1 00-1 000; positive ion linear mode detection, mass scanning range m/z 100-2000.
  • the results are shown in Fig. 6.
  • the red line represents the oil secreted by the normal sebaceous gland
  • the green line represents the oil secreted by the regenerated sebaceous gland.
  • the mass spectrometry results show that the regenerated sebaceous gland has a similar oil secretion capacity as the normal sebaceous gland.
  • the method for forming sebaceous glands of the present invention can effectively form skin tissue having sebaceous glands, and the method is simple, convenient, quick, easy to control, and easy to achieve large-scale production.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Dermatology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Transplantation (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Materials For Medical Uses (AREA)

Abstract

L'invention concerne un tissu cutané contenant une glande sébacée, le procédé de formation et l'utilisation de celui-ci, le procédé de formation de glande sébacée comprenant : la co-culture de cellules hypodermiques et de cellules souches épidermiques dans un gel.
PCT/CN2015/071777 2014-12-02 2015-01-28 Tissu cutané contenant une glande sébacée, procédé de formation et utilisation de celui-ci WO2016086527A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201410723513.5A CN104491931B (zh) 2014-12-02 2014-12-02 含有皮脂腺的皮肤组织及其形成方法和用途
CN201410723513.5 2014-12-02

Publications (1)

Publication Number Publication Date
WO2016086527A1 true WO2016086527A1 (fr) 2016-06-09

Family

ID=52933440

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2015/071777 WO2016086527A1 (fr) 2014-12-02 2015-01-28 Tissu cutané contenant une glande sébacée, procédé de formation et utilisation de celui-ci

Country Status (2)

Country Link
CN (1) CN104491931B (fr)
WO (1) WO2016086527A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109943520A (zh) * 2019-03-08 2019-06-28 北京达博威迎医药技术有限公司 汗腺细胞的分离和培养获得汗腺类器官的方法及其应用
CN113310963A (zh) * 2021-05-31 2021-08-27 四川大学华西医院 一种改进的中性粒细胞NETs免疫荧光检测方法
CN114591894A (zh) * 2022-02-28 2022-06-07 中国人民解放军总医院 一种皮肤多能前体干细胞的制备方法及其应用

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104818247B (zh) * 2015-05-18 2017-06-16 云南和泽西南生物科技有限公司 一种间充质干细胞的培养方法及用途
CN107802891A (zh) * 2017-11-09 2018-03-16 清华大学深圳研究生院 组织工程皮肤及其制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003051419A1 (fr) * 2001-12-19 2003-06-26 Henkel Kommanditgesellschaft Auf Aktien Equivalent dermique/capillaire avec papille reconstruite
CN102091352A (zh) * 2009-12-09 2011-06-15 中国人民解放军总医院第一附属医院 一种含有汗腺的组织工程皮肤模型的构建方法
CN102949752A (zh) * 2011-08-22 2013-03-06 中国人民解放军总医院第一附属医院 一种含有完全皮肤附属器的组织工程皮肤的构建方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101773688B (zh) * 2010-02-05 2013-10-23 中国人民解放军第四军医大学 一种含附属器的组织工程皮肤的制备方法
CN102172337B (zh) * 2011-02-18 2013-10-23 中国人民解放军第四军医大学 具有皮脂腺样结构的组织工程皮肤及其制备方法
US9533013B2 (en) * 2013-03-13 2017-01-03 University Of North Carolina At Chapel Hill Method of treating pancreatic and liver conditions by endoscopic-mediated (or laparoscopic-mediated) transplantation of stem cells into/onto bile duct walls of particular regions of the biliary tree

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003051419A1 (fr) * 2001-12-19 2003-06-26 Henkel Kommanditgesellschaft Auf Aktien Equivalent dermique/capillaire avec papille reconstruite
CN102091352A (zh) * 2009-12-09 2011-06-15 中国人民解放军总医院第一附属医院 一种含有汗腺的组织工程皮肤模型的构建方法
CN102949752A (zh) * 2011-08-22 2013-03-06 中国人民解放军总医院第一附属医院 一种含有完全皮肤附属器的组织工程皮肤的构建方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
FU, GANG.: "Experimental Study on Induced Differentiation of Rat Hair Follicle Bulge Cells.", SEBOCYTES IN INTRO FOURTH MILITARY MEDICAL UNIVERSITY THESIS., 31 December 2006 (2006-12-31) *
TAO, KE.: "Isolation, Cultivation of Human Fetal Sebocytes and Sweat Eccrine Gland Cells in Vitro, and the Preliminary Study on Differentiation from Human Fetal Epidermal Stem Cells to Hair Follicle", SEBACEOUS GLAND AND ECCRINE SWEAT GLAND. FOURTH MILITARY MEDICAL UNIVERSITY THESIS., 31 December 2005 (2005-12-31) *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109943520A (zh) * 2019-03-08 2019-06-28 北京达博威迎医药技术有限公司 汗腺细胞的分离和培养获得汗腺类器官的方法及其应用
CN113310963A (zh) * 2021-05-31 2021-08-27 四川大学华西医院 一种改进的中性粒细胞NETs免疫荧光检测方法
CN113310963B (zh) * 2021-05-31 2023-12-01 四川大学华西医院 一种改进的中性粒细胞NETs免疫荧光检测方法
CN114591894A (zh) * 2022-02-28 2022-06-07 中国人民解放军总医院 一种皮肤多能前体干细胞的制备方法及其应用
CN114591894B (zh) * 2022-02-28 2024-01-09 中国人民解放军总医院 一种皮肤多能前体干细胞的制备方法及其应用

Also Published As

Publication number Publication date
CN104491931B (zh) 2017-04-12
CN104491931A (zh) 2015-04-08

Similar Documents

Publication Publication Date Title
Balañá et al. Epidermal stem cells and skin tissue engineering in hair follicle regeneration
Kang et al. 3D bioprinting of a gelatin-alginate hydrogel for tissue-engineered hair follicle regeneration
JP3728750B2 (ja) 培養皮膚及びその製造方法
Schneider et al. Long-term survival and characterisation of human umbilical cord-derived mesenchymal stem cells on dermal equivalents
WO2016086527A1 (fr) Tissu cutané contenant une glande sébacée, procédé de formation et utilisation de celui-ci
JP5600671B2 (ja) 毛小嚢及びデノボ乳頭の作製方法並びにインビトロ試験及びインビボ移植のためのそれらの使用
KR20070015519A (ko) 생체 조직 시트, 그것의 제조 방법, 및 그것을 이용한 이식방법
US9592257B2 (en) Complete human skin organ generated from culture-expanded cells
US8617882B2 (en) Skin-derived precursor cells and uses thereof
CN105079783A (zh) 药物组合物及其制备方法和用途
Bannasch et al. Cultured keratinocytes in fibrin with decellularised dermis close porcine full-thickness wounds in a single step
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
US9655930B2 (en) Compositions and methods for producing reconstituted skin
Rogovaya et al. Reconstruction of rabbit urethral epithelium with skin keratinocytes
Minjuan et al. Hair Follicle Morphogenesis in the Treatment of Mouse Full‐Thickness Skin Defects Using Composite Human Acellular Amniotic Membrane and Adipose Derived Mesenchymal Stem Cells
JP2013500738A (ja) 皮膚線維芽細胞を有する細胞の支持体
EP0980270B1 (fr) Tissu de gaine dermique pour la cicatrisation
JP6839003B2 (ja) 再構成頭皮モデルおよび活性分子のスクリーニング方法
US20190328790A1 (en) Cell-embedded beads for hair regeneration and method for producing same, and kit for hair regeneration
CN109722410B (zh) 一种3d全层皮肤模型及用于其形成的培养基、制备方法
JP2020202754A (ja) 三次元培養皮膚の製造方法、及びそれにより得られる三次元培養皮膚
Flaxman Cell identification in primary cell cultures from skin
CN107174653B (zh) 一种促进毛囊再生的方法
CN103893831A (zh) 一种器官型人工皮肤、其制备方法及应用
Liu et al. Accelerating the healing of skin defects transplanted FSC-seeded tissue-engineered skin

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15865853

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15865853

Country of ref document: EP

Kind code of ref document: A1