Efficient Generation of Glucose-Responsive Beta Cells from Isolated GP2+ Human Pancreatic Progenitors

Publication date: 4 April 2017
Source:Cell Reports, Volume 19, Issue 1
Author(s): Jacqueline Ameri, Rehannah Borup, Christy Prawiro, Cyrille Ramond, Karen A. Schachter, Raphael Scharfmann, Henrik Semb
Stem cell-based therapy for type 1 diabetes would benefit from implementation of a cell purification step at the pancreatic endoderm stage. This would increase the safety of the final cell product, allow the establishment of an intermediate-stage stem cell bank, and provide a means for upscaling β cell manufacturing. Comparative gene expression analysis revealed glycoprotein 2 (GP2) as a specific cell surface marker for isolating pancreatic endoderm cells (PECs) from differentiated hESCs and human fetal pancreas. Isolated GP2⁺ PECs efficiently differentiated into glucose responsive insulin-producing cells in vitro. We found that in vitro PEC proliferation declines due to enhanced expression of the cyclin-dependent kinase (CDK) inhibitors CDKN1A and CDKN2A. However, we identified a time window when reducing CDKN1A or CDKN2A expression increased proliferation and yield of GP2⁺ PECs. Altogether, our results contribute tools and concepts toward the isolation and use of PECs as a source for the safe production of hPSC-derived β cells.

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Teaser

Ameri et al. identify GP2 as a specific marker of human pancreatic endoderm cells (PECs) and demonstrate that isolated GP2⁺ PECs generate cultures enriched in glucose-responsive insulin-producing cells. By eliminating undifferentiated hESCs, this work suggests a safer route toward manufacture of endocrine cells for future diabetes cell therapy.


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