DK2768948T3 - Fremstilling af en human beta-cellelinje fra en tidlig postnatal bugspytkirtel - Google Patents

Fremstilling af en human beta-cellelinje fra en tidlig postnatal bugspytkirtel Download PDF

Info

Publication number
DK2768948T3
DK2768948T3 DK12781298.0T DK12781298T DK2768948T3 DK 2768948 T3 DK2768948 T3 DK 2768948T3 DK 12781298 T DK12781298 T DK 12781298T DK 2768948 T3 DK2768948 T3 DK 2768948T3
Authority
DK
Denmark
Prior art keywords
cells
human
insulin
pancreatic
gene
Prior art date
Application number
DK12781298.0T
Other languages
English (en)
Inventor
Paul Czernichow
Philippe Ravassard
Raphael Scharfmann
Original Assignee
Univercell Biosolutions
Inst Nat De La Sante Et De La Rech Medicale (Inserm)
Centre Nat De La Rech Scient (C N R S )
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 Univercell Biosolutions, Inst Nat De La Sante Et De La Rech Medicale (Inserm), Centre Nat De La Rech Scient (C N R S ) filed Critical Univercell Biosolutions
Application granted granted Critical
Publication of DK2768948T3 publication Critical patent/DK2768948T3/da

Links

Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027New or modified breeds of vertebrates
    • A01K67/0271Chimeric vertebrates, e.g. comprising exogenous cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0676Pancreatic cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0676Pancreatic cells
    • C12N5/0677Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0693Tumour cells; Cancer cells
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5023Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects on expression patterns
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966Animal cells
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2207/00Modified animals
    • A01K2207/12Animals modified by administration of exogenous cells
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00Animals characterised by species
    • A01K2227/10Mammal
    • A01K2227/105Murine
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00Animals characterised by purpose
    • A01K2267/02Animal zootechnically ameliorated
    • A01K2267/025Animal producing cells or organs for transplantation
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00Animals characterised by purpose
    • A01K2267/03Animal model, e.g. for test or diseases
    • A01K2267/035Animal model for multifactorial diseases
    • A01K2267/0362Animal model for lipid/glucose metabolism, e.g. obesity, type-2 diabetes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30Hormones
    • C12N2501/33Insulin
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00Active agents used in cell culture processes, e.g. differentation
    • C12N2501/70Enzymes
    • C12N2501/73Hydrolases (EC 3.)
    • C12N2501/734Proteases (EC 3.4.)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2503/00Use of cells in diagnostics
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00Genetically modified cells
    • C12N2510/04Immortalised cells

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Environmental Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Pathology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Physics & Mathematics (AREA)
  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Toxicology (AREA)
  • Oncology (AREA)
  • Virology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Claims (14)

1. Fremgangsmåde til fremstilling af humane pankreatiske beta-celler eller humane beta-celle tumorer, omfattende trinnene: a) at dissociere neonatalt humant pankreasvæv fra et individ på mindre end 5 år med collagenase in vitro for at opnå neonatale humane pankreasceller, b) at transducere og co-transducere de neonatale humane pankreasceller opnået i trin a) med i) en lentivirusvektor der udtrykker stort T-antigen af SV40 under kontrollen af insulinpromoteren eller ii) med en lentivirusvektor der udtrykker stort T-antigen af SV40 under kontrollen af insulinpromoteren og en lentivirusvektor der udtrykker hTERT under kontrollen af insulinpromoteren, eller iii) en lentivirusvektor der udtrykker både stort T-antigen af SV40 og hTERT, c) at indføre de transducerede neonatale pankreasceller opnået i b) i nyrekapslen af et ikke-menneskeligt dyr med svær kombineret immundefekt (scid), d) at tillade de transducerede pankreasceller at udvikle insulinom-lignende strukturer, hvor de neonatale humane pankreasceller i insulinom-lignende strukturer er differentierede i insulinproducerende pankreatiske beta-celler; e) at mikrodissekere de insulinom-lignende strukturer opnået i trin d), for derved at dissociere cellerne deraf, f) at sub-transplantere cellerne opnået i trin e) ind i nyrekapslen af et nyt ikke-menneskeligt dyr med scid, g) at tillade de sub-transplanterede celler i trin f) at udvikle og regenerere insulinom-lignende strukturer, hvor de nyudviklede insulinom-lignende strukturer er beriget i insulinproducerende pankreatiske beta-celler; h) at mikrodissekere de insulinom-lignende strukturer opnået i trin g), og at dissociere og indsamle cellerne deraf, i) eventuelt, at sub-transplantere cellerne opnået i trin g) ind i nyrekapslen af et nyt ikke-menneskeligt dyr med scid, for derved at tillade yderligere berigelse og amplifikation af insulinproducerende pankreatiske beta-celler; og j) eventuelt at gentage trinnene f), g) og h), indtil den passende mængde af insulinproducerende pankreatiske beta-celler opnås.
2. Fremgangsmåden ifølge krav 1, hvor konstruktionen af lentivirusvektorerne tillader reversibel eller betinget immortalisering.
3. Fremgangsmåden ifølge et hvilket som helst af kravene 1 eller 2, hvor lentivirusvektorerne omfatter mindst et LoxP-sted og stort T-antigenet af SV40 og/eller hTERT-genet fjernes ved virkningen af Cre-rekombinasen.
4. Fremgangsmåden ifølge et hvilket som helst af kravene 1 eller 2, hvor lentivirusvektorerne omfatter mindst et FRT-sted og stort T-antigenet af SV40 og/eller hTERT-genet fjernes ved virkningen af FLP-rekombinasen.
5. Fremgangsmåden ifølge et hvilket som helst af kravene 1 eller 2, hvor lentivirusvektoren der udtrykker stort T-antigen af SV40 og lentivirusvektoren der udtrykker hTERT yderligere omfatter et LoxP- eller et FLP-sted, forudsat at de sted-specifikke rekombinationssteder er forskellige i vektorerne.
6. Fremgangsmåden ifølge et hvilket som helst af kravene 3 til 5, hvor et negativt selektionstrin udføres efter virkningen af Cre- eller FLP-rekombinasen for kun at selektere cellerne, hvor de immortaliserede gener stort T-antigen af SV40 og/eller hTERT er blevet fjernet.
7. Fremgangsmåden ifølge et hvilket som helst af kravene 3 til 5, hvor lentivirusvektorerne inkluderer mindst et negativt selektions-markørgen.
8. Fremgangsmåden ifølge krav 7, hvor det negative selektions-markørgen vælges fra gruppen udgjort af HSV-TK-genet, hypoxanthin-phosphoribosyl-transferase (HPRT)-genet, guanin-phosphoribosyl-transferase (Gpt)-genet, og cytosin-deaminase-genet.
9. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 8, hvor det ikke-menneskelige dyr med scid er en scid-mus.
10. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 9, yderligere omfattende at transducere cellerne af trin e) med en lentivirusvektor der udtrykker et antibiotisk resistensgen under kontrollen af insulinpromoteren.
11. Fremgangsmåden ifølge krav 10, hvor det antibiotiske resistensgen er et neomycin-resistensgen.
12. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 11, yderligere omfattende at indsamle de humane pankreatiske beta-celler opnået at trin j) for at danne en homogen cellepopulation og eventuelt at dyrke populationen in vitro for at etablere en human funktionel beta-cellelinje.
13. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 12, hvor collagenasen er collagenase XI.
14. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 13, yderligere omfattende et eller flere de-immortaliseringstrin, der inkluderer at fjerne stort T-antigenet af SV40, hTERT og/eller de antibiotiske resistenstransgener.
DK12781298.0T 2011-10-18 2012-10-18 Fremstilling af en human beta-cellelinje fra en tidlig postnatal bugspytkirtel DK2768948T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201161548526P 2011-10-18 2011-10-18
PCT/EP2012/070612 WO2013057164A1 (en) 2011-10-18 2012-10-18 Production of a human beta cell line from an early post natal pancreas

Publications (1)

Publication Number Publication Date
DK2768948T3 true DK2768948T3 (da) 2017-08-14

Family

ID=47143082

Family Applications (1)

Application Number Title Priority Date Filing Date
DK12781298.0T DK2768948T3 (da) 2011-10-18 2012-10-18 Fremstilling af en human beta-cellelinje fra en tidlig postnatal bugspytkirtel

Country Status (5)

Country Link
US (1) US9493743B2 (da)
EP (1) EP2768948B1 (da)
DK (1) DK2768948T3 (da)
ES (1) ES2634457T3 (da)
WO (1) WO2013057164A1 (da)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN120796168A (zh) 2013-06-11 2025-10-17 哈佛学院校长同事会 SC-β细胞以及用于产生其的组合物和方法
CN107614678B (zh) 2014-12-18 2021-04-30 哈佛学院校长同事会 干细胞来源的β细胞的产生方法及其使用方法
EP3234110B1 (en) 2014-12-18 2024-02-28 President and Fellows of Harvard College METHODS FOR GENERATING STEM CELL-DERIVED ß CELLS AND USES THEREOF
WO2016100898A1 (en) 2014-12-18 2016-06-23 President And Fellows Of Harvard College Serum-free in vitro directed differentiation protocol for generating stem cell-derived b cells and uses thereof
US11629337B2 (en) 2016-05-11 2023-04-18 Animal Cell Therapy—Act Production of a canine beta cell line from an immature pancreas
EP3710021A4 (en) 2017-11-15 2021-08-11 Semma Therapeutics, Inc. ISLAND CELL MANUFACTURING COMPOSITIONS AND METHOD OF USE
WO2020033879A1 (en) 2018-08-10 2020-02-13 Semma Therapeutics, Inc. Stem cell derived islet differentiation

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6110743A (en) * 1995-02-10 2000-08-29 The Regents Of The University Of California Development and use of human pancreatic cell lines
US7041634B2 (en) * 1995-09-27 2006-05-09 Emory University Method of inhibiting immune system destruction of transplanted viable cells
US7745216B2 (en) * 1999-02-10 2010-06-29 Curis, Inc. Methods and reagents for treating glucose metabolic disorders
EP1103615A1 (en) 1999-11-25 2001-05-30 Universite De Geneve Vectors capable of immortalizing non-dividing cells and cells immortalized with said vectors
US20050123521A1 (en) * 2001-06-21 2005-06-09 Regents Of The University Of California Immortalization of human cells by the ectopic expression of human telomerase reverse transcriptase
US20050193432A1 (en) * 2003-11-18 2005-09-01 Whitehead Institute For Biomedical Research Xenograft model of functional normal and malignant human breast tissues in rodents and methods thereof
WO2008010200A2 (en) 2006-07-20 2008-01-24 Daniel Farb Flow deflection devices and method for energy capture machines
CA2678870C (en) 2007-02-21 2017-07-11 Sarl Endocells Human pancreatic beta cell lines for diagnostic of diabetes

Also Published As

Publication number Publication date
EP2768948A1 (en) 2014-08-27
US9493743B2 (en) 2016-11-15
EP2768948B1 (en) 2017-06-07
WO2013057164A1 (en) 2013-04-25
ES2634457T3 (es) 2017-09-27
US20140302155A1 (en) 2014-10-09

Similar Documents

Publication Publication Date Title
DK2768948T3 (da) Fremstilling af en human beta-cellelinje fra en tidlig postnatal bugspytkirtel
JP5636192B2 (ja) 糖尿病の診断のためのヒト膵臓β細胞株
AU778929B2 (en) Pancreatic stem cells and their use in transplantation
US20030109036A1 (en) Method for differentiating islet precursor cells into beta cells
Scharfmann et al. Beta cells within single human islets originate from multiple progenitors
EP1456356B1 (en) Method of producing human beta cell lines
US11629337B2 (en) Production of a canine beta cell line from an immature pancreas
Ravassard et al. A new strategy to generate functional insulin-producing cell lines by somatic gene transfer into pancreatic progenitors
Demeterco et al. Gene therapy for diabetes
US20210369788A1 (en) Production of canine pancreatic islets from an immature pancreas
Bara Tissue engineering a pancreatic substitute based on recombinant intestinal endocrine cells
WO2007075956A2 (en) Methods for producing and using pancreatic endocrine cells
Hayek et al. FOR ẞ-CELL REPLACEMENT
Pais Stem cell and gene transfer-based approaches to generate insulin-producing cells
Hayek et al. Gene Therapeutic Approaches for β-Cell Replacement
AU2002351006A1 (en) Method of producing human beta cell lines