Bonfanti Paola, Nobecourt Estelle, Oshima Masaya, Albagli-Curiel Olivier, Laurysens Veerle, Stangé Geert, Sojoodi Mozhdeh, Heremans Yves, Heimberg Harry, Scharfmann Raphael
1 Diabetes Research Center, Vrije Universiteit Brussel , Brussels, Belgium .
2 INSERM, U1016, Institut Cochin, Université Paris Descartes , Sorbonne Paris Cité, Faculté de Médecine Paris, Paris, France .
Stem Cells Dev. 2015 Aug 1;24(15):1766-78. doi: 10.1089/scd.2014.0550. Epub 2015 Jun 8.
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 HNF1β) 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.
对小鼠和人类胰腺发育的比较分析可能会揭示控制器官形态发生、细胞增殖和分化等关键步骤的共同机制。更具体地说，尽管最近在从人类胚胎干细胞和诱导多能干细胞生成β细胞方面取得了进展，但了解β细胞的发育仍然是一个问题。在本研究中，我们采用了一种综合方法，包括前瞻性分离、器官培养以及对中间阶段的表征，并报告了来自人类和小鼠胎儿胰腺的细胞能够长期扩增，并在三维培养中形成中空的导管样结构。扩增后的细胞表达了一组揭示胰腺祖细胞身份的标志物（E-钙黏蛋白、PDX1、NKX6-1、SOX9和HNF1β）。Wnt激动剂R-spondin1（RSPO1）、FGF10和EGF刺激胚胎祖细胞的增殖。这种生长因子组合能够使人类胎儿胰腺祖细胞在培养中传代许多次，这一发现此前未见报道。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。我们得出结论，EGF信号的调节影响小鼠和人类胚胎胰腺祖细胞的体外扩增和分化。
