Generation of pancreatic β cells from human pluripotent stem cells (hPSCs) holds promise as a cell replacement therapy for diabetes. In this study, we establish a link between the state of the actin cytoskeleton and the expression of pancreatic transcription factors that drive pancreatic lineage specification. Bulk and single-cell RNA sequencing demonstrated that different degrees of actin polymerization biased cells toward various endodermal lineages and that conditions favoring a polymerized cytoskeleton strongly inhibited neurogenin 3-induced endocrine differentiation. Using latrunculin A to depolymerize the cytoskeleton during endocrine induction, we developed a two-dimensional differentiation protocol for generating human pluripotent stem-cell-derived β (SC-β) cells with improved in vitro and in vivo function. SC-β cells differentiated from four hPSC lines exhibited first- and second-phase dynamic glucose-stimulated insulin secretion. Transplantation of islet-sized aggregates of these cells rapidly reversed severe preexisting diabetes in mice at a rate close to that of human islets and maintained normoglycemia for at least 9 months.

从人类多能干细胞 (hPSC) 生成胰腺 β 细胞有望成为糖尿病的细胞替代疗法。在这项研究中，我们建立了肌动蛋白细胞骨架状态与驱动胰腺谱系规范的胰腺转录因子表达之间的联系。批量和单细胞 RNA 测序表明，不同程度的肌动蛋白聚合使细胞偏向各种内胚层谱系，并且有利于聚合细胞骨架的条件强烈抑制神经原 3 诱导的内分泌分化。在内分泌诱导期间使用 latrunculin A 解聚细胞骨架，我们开发了一种二维分化方案，用于生成具有改善的体外和体内功能的人多能干细胞衍生 β (SC-β) 细胞。从四个 hPSC 系分化的 SC-β 细胞表现出第一和第二阶段动态葡萄糖刺激的胰岛素分泌。移植这些细胞的胰岛大小的聚集体以接近人类胰岛的速度迅速逆转了小鼠中严重的预先存在的糖尿病，并维持正常血糖至少 9 个月。