Ex Vivo Expansion and Differentiation of Human and Mouse Fetal Pancreatic Progenitors Are Modulated by Epidermal Growth Factor
Ex Vivo Expansion and Differentiation of Human and Mouse Fetal Pancreatic Progenitors Are Modulated by Epidermal Growth Factor
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DOI：
10.1089/scd.2014.0550
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发表时间：
2015-08-01
影响因子：
4
通讯作者：
Scharfmann, Raphael
中科院分区：
文献类型：
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作者：
Bonfanti, Paola;Nobecourt, Estelle;Scharfmann, Raphael
对小鼠和人类胰腺发育的比较分析可能揭示控制器官形态发生和细胞增殖与分化等关键步骤的共同机制。更具体地说，理解β细胞发育仍然是一个问题，尽管最近的进展涉及到他们从人类胚胎和诱导多能干细胞的一代。在这项研究中，我们使用了一种综合的方法，包括前瞻性分离，器官培养和中间阶段的表征，并报告说，来自人类和小鼠胎儿胰腺的细胞可以长期扩增，并在3D培养中产生中空导管样结构。扩增的细胞表达揭示胰腺祖细胞身份的标志物（E-钙粘蛋白、PDX1、NKX6 - 1、SOX 9和HNF 1）的组合。通过Wnt激动剂R-spondin1（RSPO1）、FGF 10和EGF刺激胚胎祖细胞的增殖。这种生长因子的组合允许在培养物中维持人胎儿胰腺祖细胞多次传代，这是以前没有报道的发现。重要的是，在没有EGF的情况下，增殖减少，而内分泌分化显著增强。我们的结论是EGF信号的调制影响在体外扩增和分化的小鼠和人胚胎胰腺祖细胞。
A comparative analysis of mouse and human pancreatic development may reveal common mechanisms that control key steps as organ morphogenesis and cell proliferation and differentiation. More specifically, understanding beta cell development remains an issue, despite recent progress related to their generation from human embryonic and induced pluripotent stem cells. In this study, we use an integrated approach, including prospective isolation, organ culture, and characterization of intermediate stages, and report that cells from human and mouse fetal pancreas can be expanded in the long term and give rise to hollow duct-like structures in 3D cultures. The expanded cells express a combination of markers (E-cadherin, PDX1, NKX6-1, SOX9, and HNF1) that reveals pancreatic progenitor identity. Proliferation of embryonic progenitors was stimulated by the Wnt agonist R-spondin1 (RSPO1), FGF10, and EGF. This combination of growth factors allowed maintaining human fetal pancreatic progenitors in culture for many passages, a finding not reported previously. Importantly, in the absence of EGF, proliferation was reduced, while endocrine differentiation was significantly enhanced. We conclude that modulation of EGF signaling affects in vitro expansion and differentiation of progenitors from embryonic pancreas of both mice and man.