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

作者：Xisheng Li, Yi Yang, Vicken W. Chan, Kam Tong Leung, Xiao‐Bing Zhang, Alan S.L. Wong, Charing C. N. Chong, Chi Chiu Wang, Manching Ku, Kathy O. Lui · 发表于：Stem Cell Reports · 年份：2020 · DOI：10.1016/j.stemcr.2020.09.009 · 被引用次数：49 · 研究领域：Pancreatic function and diabetes、Diabetes and associated disorders

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