Department of Chemical Pathology; Li Ka Shing Institute of Health Sciences, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China.
Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China.
Stem Cell Reports. 2020 Nov 10;15(5):1111-1126. doi: 10.1016/j.stemcr.2020.09.009. Epub 2020 Oct 22.
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 NKX6.1MAFAC-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 CD9C-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.
迄今为止，尚不清楚是否存在用于从人多能干细胞 (hPSC) 中纯化葡萄糖反应性胰岛样 β 样细胞的特定细胞表面标志物。在寻找这个标志物的过程中，我们开发了一种从人胚胎干细胞中高效分化β样细胞的方案。我们进行了单细胞 RNA 测序，发现 CD9 是β样细胞的阴性细胞表面标志物，因为大多数 INS 细胞都是 CD9。我们针对 CD9 进行了β样细胞的自发形成胰岛样类器官的纯化，发现 CD9 群体中的 NKX6.1+MAFAC-PEPTIDE+β样细胞明显更多。CD9 细胞的葡萄糖反应性也优于 CD9 细胞。在人类中，我们观察到胎儿胰岛中 CD9C+PEPTIDE+β 细胞比成人尸体胰岛中更多，而且 CD9 胎儿 β 细胞中的 Ki67 增殖细胞更多。综上所述，我们的实验表明 CD9 是人多能干细胞分化的葡萄糖反应性 β 样细胞的阴性富集的细胞表面标志物。
