Long-term feeder-free culture of human pancreatic progenitors on fibronectin or matrix-free polymer potentiates β cell differentiation
Open accessAbstract
With the aim of producing β cells for replacement therapies to treat diabetes, several protocols have been developed to differentiate human pluripotent stem cells to β cells via pancreatic progenitors. While in vivo pancreatic progenitors expand throughout development, the in vitro protocols have been designed to make these cells progress as fast as possible to β cells. Here, we report on a protocol enabling a long-term expansion of human pancreatic progenitors in a defined medium on fibronectin, in the absence of feeder layers. Moreover, through a screening of a polymer library we identify a polymer that can replace fibronectin. Our experiments, comparing expanded progenitors to directly differentiated progenitors, show that the expanded progenitors differentiate more efficiently into glucose-responsive β cells and produce fewer glucagon-expressing cells. The ability to expand progenitors under defined conditions and cryopreserve them will provide flexibility in research and therapeutic production.
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Defined culture media containing either SB431542 or I-BET151 promoted the expansion of PDX1+/NKX6-1+ human pancreatic progenitors.
“In particular, it was recently reported that defined culture media containing either the TGF-β pathway inhibitor SB431542183 or I-BET151,184 an inhibitor of the acetyl-lysine bromodomain-containing proteins, promoted the expansion of PDX1+/NKX6-1+ human pancreatic progenitors.”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [183]
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- Authors
- Nakamura, A., Wong, Y. F., Venturato, A., Michaut, M., Venkateswaran, S., Santra, M., Gonçalves, C., Larsen, M., Leuschner, M., Kim, Y. H., Brickman, J., Bradley, M., & Grapin-Botton, A.
- Affiliations
- University of Copenhagen (DK)
- University of Edinburgh (GB)
- Max Planck Institute of Molecular Cell Biology and Genetics (DE)
- ORCID
- Venue
- Stem Cell Reports
- Publisher
- Elsevier BV
- Published
- 2022-05
- Volume
- 17
- Issue
- 5
- Pages
- 1215-1228
- ISSN
- 2213-6711 (ISSN-L)
- PMC
- PMC9133655
- OpenAlex
- W4224281399
- Cited by
- 20
- References
- 34
- Citations by year
- 2025: 4 · 2024: 5 · 2023: 10 · 2022: 1
- FWCI
- 5.73
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Stem Cell Reports (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by-nc-nd)
- Queen Mary Research Online (Queen Mary University of London) (submittedVersion, cc-by-nc-nd)
- Site cant be reached (submittedVersion, cc-by-nc-nd)
- Edinburgh Research Explorer (University of Edinburgh) (submittedVersion, cc-by-nc-nd)
- PuSH - Publication Server of Helmholtz Zentrum München (submittedVersion, cc-by-nc-nd)
- Funders
- European Commission Seventh Framework Programme for Research and Technological Development Health
- Max Planck Society
- Novo Nordisk Fonden (NNF17CC0027852, NNF21CC0073729)
- Topics
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
- Keywords
- feeder-free culture, fibronectin, beta cell differentiation, glucagon, pluripotent stem cells, cryopreservation, chemically defined medium
- MeSH terms
- Cell Differentiation, Fibronectins, Humans, Pancreas, Polymers, Pluripotent Stem Cells, Insulin-Secreting Cells
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