Single-Cell RNA-Seq Reveals that CD9 Is a Negative Marker of Glucose-Responsive Pancreatic ?-like Cells Derived from Human Pluripotent Stem Cells.

ABSTRACT: To date, it remains unclear if there are specific cell-surface markers for purifying glucose-responsive pancreatic ?-like cells derived from human pluripotent stem cells (hPSCs). In searching for this, we generated an efficient protocol for differentiating ?-like cells from human embryonic stem cells. We performed single-cell RNA sequencing and found that CD9 is a negative cell-surface marker of ?-like cells, as most INS+ cells are CD9-. We purified ?-like cells for spontaneous formation of islet-like organoids against CD9, and found significantly more NKX6.1+MAFA+C-PEPTIDE+ ?-like cells in the CD9- than in the CD9+ population. CD9- cells also demonstrate better glucose responsiveness than CD9+ cells. In humans, we observe more CD9+C-PEPTIDE+ ? cells in the fetal than in the adult cadaveric islets and more Ki67+ proliferating cells among CD9+ fetal ? cells. Taken together, our experiments show that CD9 is a cell-surface marker for negative enrichment of glucose-responsive ?-like cells differentiated from hPSCs.

SUBMITTER: Li X 

PROVIDER: S-EPMC7663789 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature