Abstract
PDX1+/NKX6-1+ pancreatic progenitors (PPs) give rise to endocrine cells both in vitro and in vivo. This cell population can be successfully differentiated from human pluripotent stem cells (hPSCs) and hold the potential to generate an unlimited supply of β cells for diabetes treatment. However, the efficiency of PP generation in vitro is highly variable, negatively impacting reproducibility and validation of in vitro and in vivo studies, and consequently, translation to the clinic. Here, we report the use of a proteomics approach to phenotypically characterize hPSC-derived PPs and distinguish these cells from non-PP populations during differentiation. Our analysis identifies the pancreatic secretory granule membrane major glycoprotein 2 (GP2) as a PP-specific cell surface marker. Remarkably, GP2 is co-expressed with NKX6-1 and PTF1A in human developing pancreata, indicating that it marks the multipotent pancreatic progenitors in vivo. Finally, we show that isolated hPSC-derived GP2+ cells generate β-like cells (C-PEPTIDE+/NKX6-1+) more efficiently compared to GP2− and unsorted populations, underlining the potential therapeutic applications of GP2.
Highlights
PDX1+/NKX6-1+ pancreatic progenitors (PPs) give rise to endocrine cells both in vitro and in vivo
To provide a safer cell population for therapeutic purposes and obviate the risk of contamination from undifferentiated human pluripotent stem cells (hPSCs) and/or other germ layer derivatives, we set out to identify specific cell surface markers that will allow for quantification and enrichment of hPSC-derived PPs
Driven by the proteomic results showing that glycoprotein 2 (GP2) is highly enriched in the human embryonic stem cell (hESC)-derived pancreatic progenitor cells, we set out to investigate the expression of GP2 in the human postnatal pancreas and revealed the existence of a subset of cells coexpressing PTF1A and NKX6-1 that localize to the epithelial tips in the human pancreas at birth (Fig. 2g)
Summary
PDX1+/NKX6-1+ pancreatic progenitors (PPs) give rise to endocrine cells both in vitro and in vivo. This cell population can be successfully differentiated from human pluripotent stem cells (hPSCs) and hold the potential to generate an unlimited supply of β cells for diabetes treatment. We identify the pancreatic secretory granule membrane major glycoprotein 2 (GP2) as a specific cell surface marker of hESCderived PPs. We confirm expression of GP2 in sections of human developing pancreas, marking the NKX6-1+/PTF1A+ putative human multipotent progenitors. Through the use of automated magnetic cell isolation, GP2 enrichment could be used for large-scale production of hESC-derived pancreatic progenitor cells for clinical and pharmaceutical purposes
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