Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells.
Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells.
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DOI：
10.1016/j.stemcr.2017.05.019
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发表时间：
2017-06-06
影响因子：
5.9
通讯作者：
Dunn NR
中科院分区：
文献类型：
--
作者：
Trott J;Tan EK;Ong S;Titmarsh DM;Denil SLIJ;Giam M;Wong CK;Wang J;Shboul M;Eio M;Cooper-White J;Cool SM;Rancati G;Stanton LW;Reversade B;Dunn NR
多能干细胞已被提议作为研究和治疗糖尿病的胰腺β细胞的无限来源。然而，用于产生功能性辟田胞的长的多步骤分化方案不可避免地表现出相当大的可变性，特别是当应用于来自不同遗传背景的多能细胞时。我们已经开发了支持人多能胰腺祖细胞长期自我更新的培养条件，这些祖细胞在发育上更接近成人胰腺的特化细胞。这些培养的胰腺祖细胞（cPP）表达关键的胰腺转录因子，包括PDX1和SOX 9，并表现出与其体内对应物密切相关的转录组。在暴露于分化线索后，cPP细胞产生胰腺内分泌、腺泡和导管谱系，表明多谱系潜能。此外，cPP细胞在体外和体内产生胰岛素+ β样细胞，表明它们提供了多能细胞的方便替代物，作为用于建模胰腺发育和糖尿病的成体细胞类型的来源。在3T3细胞上培养能够实现人胰腺祖细胞的长期自我更新增殖需要EGF、FGF 10、视黄酸以及Notch和TGF-β的抑制培养的祖细胞上调有丝分裂和端粒维持所需的基因胰管和β样细胞在体外和体内产生。Trott及其同事描述了使来自人类多能干细胞的胰腺祖细胞能够长期自我更新的条件。这些培养的胰腺祖细胞可以扩增至少20代，并且能够在体外和体内分化成多种胰腺谱系，包括β样细胞。
Pluripotent stem cells have been proposed as an unlimited source of pancreatic β cells for studying and treating diabetes. However, the long, multi-step differentiation protocols used to generate functional β cells inevitably exhibit considerable variability, particularly when applied to pluripotent cells from diverse genetic backgrounds. We have developed culture conditions that support long-term self-renewal of human multipotent pancreatic progenitors, which are developmentally more proximal to the specialized cells of the adult pancreas. These cultured pancreatic progenitor (cPP) cells express key pancreatic transcription factors, including PDX1 and SOX9, and exhibit transcriptomes closely related to their in vivo counterparts. Upon exposure to differentiation cues, cPP cells give rise to pancreatic endocrine, acinar, and ductal lineages, indicating multilineage potency. Furthermore, cPP cells generate insulin+ β-like cells in vitro and in vivo, suggesting that they offer a convenient alternative to pluripotent cells as a source of adult cell types for modeling pancreatic development and diabetes. Culture on 3T3 cells enables long-term self-renewal of human pancreatic progenitors Proliferation requires EGF, FGF10, retinoic acid, and inhibition of Notch and TGF-β Cultured progenitors upregulate genes required for mitosis and telomere maintenance Pancreatic duct and β-like cells are generated in vitro and in vivo In this article, Trott and colleagues describe conditions that enable long-term self-renewal of pancreatic progenitors derived from human pluripotent stem cells. These cultured pancreatic progenitors can be expanded for at least 20 passages and are capable of differentiation into multiple pancreatic lineages, including β-like cells, both in vitro and in vivo.