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Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids

Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids
Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids

Background: Pancreatic organoid systems have recently been described for the in vitro culture of pancreatic ductal cells from mouse and human. Mouse pancreatic organoids exhibit unlimited expansion potential, while previously reported human pancreas organoid (hPO) cultures do not expand efficiently long-term in a chemically defined, serum-free medium. We sought to generate a 3D culture system for long-term expansion of human pancreas ductal cells as hPOs to serve as the basis for studies of human pancreas ductal epithelium, exocrine pancreatic diseases and the development of a genomically stable replacement cell therapy for diabetes mellitus. Results: Our chemically defined, serum-free, human pancreas organoid culture medium supports the generation and expansion of hPOs with high efficiency from both fresh and cryopreserved primary tissue. hPOs can be expanded from a single cell, enabling their genetic manipulation and generation of clonal cultures. hPOs expanded for months in vitro maintain their ductal morphology, biomarker expression and chromosomal integrity. Xenografts of hPOs survive long-term in vivo when transplanted into the pancreas of immunodeficient mice. Notably, mouse orthotopic transplants show no signs of tumorigenicity. Crucially, our medium also supports the establishment and expansion of hPOs in a chemically defined, modifiable and scalable, biomimetic hydrogel. Conclusions: hPOs can be expanded long-term, from both fresh and cryopreserved human pancreas tissue in a chemically defined, serum-free medium with no detectable tumorigenicity. hPOs can be clonally expanded, genetically manipulated and are amenable to culture in a chemically defined hydrogel. hPOs therefore represent an abundant source of pancreas ductal cells that retain the characteristics of the tissue-of-origin, which opens up avenues for modelling diseases of the ductal epithelium and increasing understanding of human pancreas exocrine biology as well as for potentially producing insulin-secreting cells for the treatment of diabetes.

Chemically defined hydrogel, Genetic stability, In vivo safety, Organoid, Pancreas, Primary cultures
1471-213X
Georgakopoulos, Nikitas
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Prior, Nicole
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Angres, Brigitte
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Mastrogiovanni, Gianmarco
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Cagan, Alex
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Harrison, Daisy
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Hindley, Christopher J.
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Arnes-Benito, Robert
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Liau, Siong-Seng
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Curd, Abbie
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Ivory, Natasha
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Simons, Benjamin D.
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Martincorena, Inigo
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Wurst, Helmut
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Saeb-Parsy, Kourosh
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Huch, Meritxell
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Georgakopoulos, Nikitas
3beeb460-8761-4828-bcb1-29a9fbfcbaa7
Prior, Nicole
27851f03-6041-4409-a15c-a2a164b883e0
Angres, Brigitte
b0d4add0-a9e8-48ae-97bc-a614120449db
Mastrogiovanni, Gianmarco
8a9ef419-8ff8-48b7-a2f1-5268fc3273a9
Cagan, Alex
dbf55097-3e4d-4775-8ff1-a459523bed18
Harrison, Daisy
9e0499e9-2c71-4fcc-9a1d-920a861d727e
Hindley, Christopher J.
3f009c77-55b7-4810-b741-6a67618e1406
Arnes-Benito, Robert
a15d8267-9570-4411-8ef1-7c4c465bb499
Liau, Siong-Seng
971a914a-1500-43f4-a94a-ecd6e2db66b5
Curd, Abbie
c2e9dee6-53d6-42f7-9964-a009b98fc2c0
Ivory, Natasha
6254e3ac-ebc6-4b79-9fae-a150cc865825
Simons, Benjamin D.
02e0ea52-9b7f-4b80-a856-a22cc1991ba3
Martincorena, Inigo
5f447626-1b62-4d63-b14b-027299b8f90f
Wurst, Helmut
77843d56-5e83-470e-a580-3441f46ec39b
Saeb-Parsy, Kourosh
598a9eb8-f4b0-45e4-ac27-679cff7d2919
Huch, Meritxell
defdc21f-6ca2-4a97-ab38-436adce9d37c

Georgakopoulos, Nikitas, Prior, Nicole, Angres, Brigitte, Mastrogiovanni, Gianmarco, Cagan, Alex, Harrison, Daisy, Hindley, Christopher J., Arnes-Benito, Robert, Liau, Siong-Seng, Curd, Abbie, Ivory, Natasha, Simons, Benjamin D., Martincorena, Inigo, Wurst, Helmut, Saeb-Parsy, Kourosh and Huch, Meritxell (2020) Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids. BMC Developmental Biology, 20 (1), [4]. (doi:10.1186/s12861-020-0209-5).

Record type: Article

Abstract

Background: Pancreatic organoid systems have recently been described for the in vitro culture of pancreatic ductal cells from mouse and human. Mouse pancreatic organoids exhibit unlimited expansion potential, while previously reported human pancreas organoid (hPO) cultures do not expand efficiently long-term in a chemically defined, serum-free medium. We sought to generate a 3D culture system for long-term expansion of human pancreas ductal cells as hPOs to serve as the basis for studies of human pancreas ductal epithelium, exocrine pancreatic diseases and the development of a genomically stable replacement cell therapy for diabetes mellitus. Results: Our chemically defined, serum-free, human pancreas organoid culture medium supports the generation and expansion of hPOs with high efficiency from both fresh and cryopreserved primary tissue. hPOs can be expanded from a single cell, enabling their genetic manipulation and generation of clonal cultures. hPOs expanded for months in vitro maintain their ductal morphology, biomarker expression and chromosomal integrity. Xenografts of hPOs survive long-term in vivo when transplanted into the pancreas of immunodeficient mice. Notably, mouse orthotopic transplants show no signs of tumorigenicity. Crucially, our medium also supports the establishment and expansion of hPOs in a chemically defined, modifiable and scalable, biomimetic hydrogel. Conclusions: hPOs can be expanded long-term, from both fresh and cryopreserved human pancreas tissue in a chemically defined, serum-free medium with no detectable tumorigenicity. hPOs can be clonally expanded, genetically manipulated and are amenable to culture in a chemically defined hydrogel. hPOs therefore represent an abundant source of pancreas ductal cells that retain the characteristics of the tissue-of-origin, which opens up avenues for modelling diseases of the ductal epithelium and increasing understanding of human pancreas exocrine biology as well as for potentially producing insulin-secreting cells for the treatment of diabetes.

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Accepted/In Press date: 3 February 2020
e-pub ahead of print date: 26 February 2020
Published date: 26 February 2020
Keywords: Chemically defined hydrogel, Genetic stability, In vivo safety, Organoid, Pancreas, Primary cultures

Identifiers

Local EPrints ID: 441626
URI: http://eprints.soton.ac.uk/id/eprint/441626
ISSN: 1471-213X
PURE UUID: 0fbb7020-253a-453a-af64-f8573306cfc5
ORCID for Nicole Prior: ORCID iD orcid.org/0000-0003-2856-7052

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Date deposited: 22 Jun 2020 16:30
Last modified: 26 Nov 2021 03:22

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Contributors

Author: Nikitas Georgakopoulos
Author: Nicole Prior ORCID iD
Author: Brigitte Angres
Author: Gianmarco Mastrogiovanni
Author: Alex Cagan
Author: Daisy Harrison
Author: Christopher J. Hindley
Author: Robert Arnes-Benito
Author: Siong-Seng Liau
Author: Abbie Curd
Author: Natasha Ivory
Author: Benjamin D. Simons
Author: Inigo Martincorena
Author: Helmut Wurst
Author: Kourosh Saeb-Parsy
Author: Meritxell Huch

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