Long-term feeder-free culture of human pancreatic progenitors on fibronectin or matrix-free polymer potentiates 13 cell differentiation

With the aim of producing 13 cells for replacement therapies to treat diabetes, several protocols have been developed to differentiate human pluripotent stem cells to 13 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 13 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 13 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.

Automated summary

In this study, a protocol for long-term expansion of human pancreatic progenitors in a defined medium was developed, and a polymer that can replace fibronectin was identified. The expanded progenitors showed more efficient differentiation into glucose-responsive 13 cells and produced fewer glucagon-expressing cells compared to directly differentiated progenitors. This method provides flexibility in research and therapeutic production by allowing the expansion and cryopreservation of progenitors under defined conditions.