Long-term feeder-free culture of human pancreatic progenitors on fibronectin or matrix-free polymer potentiates β cell differentiation

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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.

OriginalsprogEngelsk
TidsskriftStem Cell Reports
Vol/bind17
Udgave nummer5
Sider (fra-til)1215-1228
Antal sider14
ISSN2213-6711
DOI
StatusUdgivet - 2022

Bibliografisk note

Funding Information:
The Novo Nordisk Foundation Center for Stem Cell Biology is supported by Novo Nordisk Foundation , Denmark grant NNF17CC0027852 , and the Novo Nordisk Foundation Center for Stem Cell Medicine is supported by Novo Nordisk Foundation , Denmark grant NNF21CC0073729 . We thank Helen Neil and the DanStem Genomics Platform and the facilities of Stem Cell Culture, Flow Cytometry, and Imaging at DanStem, as well as the facilities of Organoid and Stem Cell and Cell Technologies at MPI-CBG, and their staff for technical expertise, support, and the use of instruments. Data processing and analysis were performed using the DeiC National Life Science Supercomputer at DTU ( www.computerome.dk ).

Publisher Copyright:
© 2022 The Authors

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