Gut stem cells are accessible by biopsy and propagate robustly in culture, offering an invaluable resource for autologous cell therapies. Insulin-producing cells can be induced in mouse gut, but it has not been possible to generate abundant and durable insulin-secreting cells from human gut tissues to evaluate their potential as a cell therapy for diabetes. Here we describe a protocol to differentiate cultured human gastric stem cells into pancreatic islet-like organoids containing gastric insulin-secreting (GINS) cells that resemble β-cells in molecular hallmarks and function. Sequential activation of the inducing factors NGN3 and PDX1-MAFA led human gastric stem cells onto a distinctive differentiation path, including a SOX4^High endocrine and Galanin^High GINS precursor, before adopting β-cell identity, at efficiencies close to 70%. GINS organoids acquired glucose-stimulated insulin secretion in 10 days and restored glucose homeostasis for over 100 days in diabetic mice after transplantation, providing proof of concept for a promising approach to treat diabetes.

肠道干细胞可通过活检获得，并在培养物中强劲繁殖，为自体细胞疗法提供宝贵的资源。可以在小鼠肠道中诱导产生胰岛素的细胞，但不可能从人类肠道组织中产生丰富且持久的胰岛素分泌细胞来评估其作为糖尿病细胞疗法的潜力。在这里，我们描述了一种将培养的人胃干细胞分化为胰岛样器官的方案，其中含有胃胰岛素分泌（GINS）细胞，其分子特征和功能类似于β细胞。诱导因子 NGN3 和 PDX1-MAFA 的顺序激活导致人胃干细胞走上独特的分化途径，包括 SOX4^高内分泌和甘丙肽^高GINS 前体，然后采用 β 细胞身份，效率接近 70%。GINS 类器官在移植后 10 天内获得了葡萄糖刺激的胰岛素分泌，并在糖尿病小鼠中恢复了 100 多天的葡萄糖稳态，这为治疗糖尿病的有前途的方法提供了概念证明。