DK3072535T3 - Fremgangsmåde til rekonstruktion af hud - Google Patents

Fremgangsmåde til rekonstruktion af hud Download PDF

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Publication number
DK3072535T3
DK3072535T3 DK15305441.6T DK15305441T DK3072535T3 DK 3072535 T3 DK3072535 T3 DK 3072535T3 DK 15305441 T DK15305441 T DK 15305441T DK 3072535 T3 DK3072535 T3 DK 3072535T3
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Denmark
Prior art keywords
skin
médium
culture
substitute
fibroblasts
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DK15305441.6T
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English (en)
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DK3072535T5 (da
Inventor
Vincent Casoli
Muriel Cario-André
Jean-Christophe Lepivert
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Univ Bordeaux
Inst Nat De La Santé Et De La Rech Médicale (Inserm)
Centre Hospitalier Univ Bordeaux
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
    • 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
    • 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
    • A61L27/36—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
    • 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
    • A61L27/3813—Epithelial cells, e.g. keratinocytes, urothelial 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
    • A61L27/50—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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C12N5/0697—Artificial constructs associating cells of different lineages, e.g. tissue equivalents
    • C12N5/0698—Skin equivalents
    • 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
    • C12N2500/32—Amino acids
    • 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
    • C12N2501/90—Polysaccharides
    • C12N2501/905—Hyaluronic acid
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/09—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells
    • C12N2502/091—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells melanocytes
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/09—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells
    • C12N2502/094—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells keratinocytes
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
    • C12N2502/1323—Adult fibroblasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/50—Proteins
    • C12N2533/54—Collagen; Gelatin

Claims (14)

  1. FREMGANGSMÁDE TIL REKONSTRUKTION AF HUD
    1. Fremgangsmáde til fremstilling af en hudsubstitut omfattende f0lgende trin: a. dyrkning af fibroblaster i et fibroblastdyrkningsmedium MI; b. podning af en matrix omfattende collagen med fibroblaster opnáet fra trin a; c. dyrkning af indpodede fibroblaster i en matrix omfattende collagen i et fibroblastdyrkningsmedium M2, hvilken matrix og de dyrkede fibroblaster danner en dermissubstitut; d. dyrkning af melanocytter i et melanocytdyrkningsmedium M3; e. dyrkning af keratinocytter i et keratinocytdyrkningsmedium M4 ; f. blanding af melanocytter opnáet i trin d med keratinocytter opnáet i trin e; g. podning af dermissubstitutten opnáet i trin c med blandingen opnáet i trin f; h. dyrkning af dermissubstitut indpodet i trin g i et hudkulturmedium M5, sáledes at hudsubstitutten dannes. hvori mediet M2 omfatter ascorbinsyre eller en ascorbat, mediet M5 omfatter hyaluronsyre eller en hyaluronat eller et derivat af disse produkter, og podning i trin g udfores med et (keratinocytter + melanocytter)/fibroblaster-forhold fra 9 og 19.
  2. 2. Fremgangsmáde ifolge krav 1, hvori mediet M5 omfatter ascorbinsyre eller en ascorbat.
  3. 3. Fremgangsmáde ifolge et hvilket som helst af kravene 1 til 2, hvori blandingen af melanocytter og keratinocytter i trin f foretages med et melanocytter / keratinocytter-forhold fra 1/20 og 1/15.
  4. 4. Fremgangsmáde if0lge et hvilket som helst af de foregáende krav, hvori podning udfort i trin b udfpres ved en densitet pá mellem 20.000 og 50.000 fibroblaster / cm2 af overfladearealet af den matrixen omfattende collagen.
  5. 5. Fremgangsmáde ifolge et hvilket som helst af de foregáende krav, hvori trin c. omfatter et fprste dyrkningstrin c' i 18 til 28 timer i tilstedevasrelse af et fibroblastdyrkningsmedium M21 omfattende hverken ascorbinsyre eller ascorbat, et andet dyrkningstrin c" pá mindst 2 dage i tilstedevaerelse af et fibroblastdyrkningsmedium M22 omfattende ascorbinsyre eller en ascorbat.
  6. 6. Fremgangsmáde ifolge et hvilket som helst af de foregáende krav, hvori trin h. omfatter: - et forste 6- til 24-timers dyrkningstrin h.' i tilstedev*relse af et dyrkningsmedium M51 omfattende hverken hyaluronsyre eller hyaluronat eller ascorbinsyre eller ascorbat, - et andet dyrkningstrin h." pá mindst 2 dage i tilstedevasrelse af et dyrkningsmedium M52 omfattende hyaluronsyre eller en hyaluronat og - et tredje dyrkningstrin h.'" pá mindst 2 dage i et médium M53 omfattende hyaluronsyre eller en hyaluronat og ascorbinsyre eller en ascorbat.
  7. 7. Hudsubstitut opnáet ved implementering af fremgangsmáden defineret ved et hvilket som helst af de foregáende krav.
  8. 8. Hudsubstitut ifolge krav 7, hvori fibroblasterne er autologe fra et individ under forberedelse til en graft af hudsubstitutten pá individet.
  9. 9. Hudsubstitut ifplge krav 8, hvori fibroblasterne, melanocytterne og keratinocytterne er autologe fra et individ.
  10. 10. Graft udgjort af en hudsubstitut if0lge krav 8 eller 9.
  11. 11. Graft if0lge krav 10 til anvendelse som en behandlingsmetode for en hudlidelse og/eller et tab af hudsubstans.
  12. 12. Graft til anvendelse ifolge krav 11, hvori hudlidelsen og/eller tabet af hudsubstans er udvalgt fra gruppen bestáende af en f orbrasnding, en helingsdefekt, et kronisk sár, en pigmentf orstyrrelse, at haemangiom og en hudcancer.
  13. 13. Kit til anvendelse af fremgangsmáden ifolge et hvilket som helst af kravene 1 til 6 omfattende et fibroblastdyrkningsmedium MI, en matrix omfattende collagen, et fibroblastdyrkningsmedium M2 i matrixen omfattende collagen, et melanocytdyrkningsmedium M3, et keratinocytdyrkningsmedium M4 og et huddyrkningsmedium M5, hvori mediet M2 omfatter ascorbinsyre eller en ascorbat, og mediet M5 omfatter hyaluronsyre eller en hyaluronat.
  14. 14. Kit ifolge krav 13, hvori medierne MI til M5 uafhaengigt er medier af klinisk kvalitet.
DK15305441.6T 2015-03-26 2015-03-26 Fremgangsmåde til rekonstruktion af hud DK3072535T5 (da)

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Application Number Priority Date Filing Date Title
EP15305441.6A EP3072535B1 (fr) 2015-03-26 2015-03-26 Procédé de reconstruction de peau

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DK3072535T3 true DK3072535T3 (da) 2017-05-22
DK3072535T5 DK3072535T5 (da) 2017-06-06

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DK16713834.6T DK3274005T3 (da) 2015-03-26 2016-03-25 Fremgangsmåde til rekonstruktion af hud

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US (1) US11643639B2 (da)
EP (2) EP3072535B1 (da)
JP (1) JP6729880B2 (da)
CN (1) CN107683148B (da)
AU (1) AU2016238760B2 (da)
BR (1) BR112017020013B1 (da)
CA (1) CA2977725A1 (da)
DK (2) DK3072535T5 (da)
ES (2) ES2625768T3 (da)
WO (1) WO2016151133A1 (da)

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Publication number Priority date Publication date Assignee Title
KR102540981B1 (ko) * 2017-06-30 2023-06-08 (주)아모레퍼시픽 인공피부 제조방법 및 인공피부
CN111450321A (zh) * 2019-01-18 2020-07-28 北京华弗生物科技有限公司 人工皮肤替代物及其无支架自组装构建方法和用途
KR102460180B1 (ko) * 2020-11-27 2022-10-27 숭실대학교산학협력단 연령별 구조 및 물성을 갖는 인공피부, 및 이의 제조방법
CN115627256B (zh) * 2022-12-02 2023-03-07 上海尚瑞生物医药科技有限公司 毛囊细胞构成的多层组织工程皮肤及其制备方法与应用

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Publication number Publication date
EP3072535B1 (fr) 2017-03-01
JP2018513724A (ja) 2018-05-31
AU2016238760A1 (en) 2017-09-14
EP3274005B1 (fr) 2019-12-04
ES2625768T3 (es) 2017-07-20
ES2772700T3 (es) 2020-07-08
WO2016151133A1 (fr) 2016-09-29
CA2977725A1 (fr) 2016-09-29
JP6729880B2 (ja) 2020-07-29
EP3072535A1 (fr) 2016-09-28
AU2016238760B2 (en) 2019-11-14
DK3072535T5 (da) 2017-06-06
US11643639B2 (en) 2023-05-09
CN107683148B (zh) 2021-05-04
US20180112188A1 (en) 2018-04-26
BR112017020013B1 (pt) 2021-06-22
CN107683148A (zh) 2018-02-09
EP3274005A1 (fr) 2018-01-31
DK3274005T3 (da) 2020-02-24
BR112017020013A2 (pt) 2018-06-19

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