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.

迄今为止，尚不清楚是否存在用于纯化源自人类多能干细胞 (hPSC) 的葡萄糖反应性胰腺 β 样细胞的特定细胞表面标志物。在寻找这一点时，我们生成了一个有效的方案，用于从人类胚胎干细胞中区分 β 样细胞。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 ⁺细胞是 CD9 ^-。我们纯化了 β 样细胞以自发形成针对 CD9 的胰岛样类器官，并发现CD9 ^-中的NKX6.1 ⁺ MAFA ⁺ C-肽⁺ β 样细胞明显多于 CD9 ⁺群体。CD9 ^-细胞还表现出比 CD9 ⁺细胞更好的葡萄糖反应性。在人类中，我们观察到胎儿中的CD9 ⁺ C-PEPTIDE ⁺ β 细胞多于成人尸体胰岛，而CD9 ⁺胎儿 β 细胞中的Ki67 ⁺增殖细胞更多。总之，我们的实验表明 CD9 是一种细胞表面标记，用于从 hPSC 分化的葡萄糖反应性 β 样细胞的负富集。