Controlled induction of human pancreatic progenitors produces functional beta-like cells in vitro
Controlled induction of human pancreatic progenitors produces functional beta-like cells in vitro
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DOI：
10.15252/embj.201591058
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发表时间：
2015-07-02
期刊：
影响因子：
11.4
通讯作者：
Hebrok, Matthias
中科院分区：
文献类型：
--
作者：
Russ, Holger A.;Parent, Audrey V.;Hebrok, Matthias
将人多能干细胞定向分化为功能性胰岛素产生β样细胞为糖尿病患者的细胞替代疗法带来了巨大的希望。这种方法还提供了独特的机会，研究人类β细胞发育和功能的其他方面。在这里，我们表明，目前的胰腺祖细胞分化方案促进早熟内分泌承诺，最终导致非功能性多激素细胞的产生。在胰腺特化过程中省略常用的BMP抑制剂可以防止早熟的内分泌形成，而用维甲酸和EGF/KGF联合治疗可以分别有效地产生PDX1（+）和随后的PDX1（+）/NKX6.1（+）胰腺祖细胞群。PDX1（+）/NKX6.1（+）祖细胞中内分泌分化的精确时间激活在体外产生葡萄糖响应性β样细胞，其表现出真正的人类β细胞的关键特征，在短期移植后保持功能，并降低糖尿病小鼠的血糖水平。因此，我们的简化和可扩展的系统准确地概括了人类胰腺发育的关键步骤，并提供了功能性人类β样细胞的快速和可重复的供应。
Directed differentiation of human pluripotent stem cells into functional insulin-producing beta-like cells holds great promise for cell replacement therapy for patients suffering from diabetes. This approach also offers the unique opportunity to study otherwise inaccessible aspects of human beta cell development and function in vitro. Here, we show that current pancreatic progenitor differentiation protocols promote precocious endocrine commitment, ultimately resulting in the generation of non-functional polyhormonal cells. Omission of commonly used BMP inhibitors during pancreatic specification prevents precocious endocrine formation while treatment with retinoic acid followed by combined EGF/KGF efficiently generates both PDX1(+) and subsequent PDX1(+)/NKX6.1(+) pancreatic progenitor populations, respectively. Precise temporal activation of endocrine differentiation in PDX1(+)/NKX6.1(+) progenitors produces glucose-responsive beta-like cells in vitro that exhibit key features of bona fide human beta cells, remain functional after short-term transplantation, and reduce blood glucose levels in diabetic mice. Thus, our simplified and scalable system accurately recapitulates key steps of human pancreas development and provides a fast and reproducible supply of functional human beta-like cells.