Morrison Gillian M, Oikonomopoulou Ifigenia, Migueles Rosa Portero, Soneji Shamit, Livigni Alessandra, Enver Tariq, Brickman Joshua M
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JQ, UK.
Cell Stem Cell. 2008 Oct 9;3(4):402-15. doi: 10.1016/j.stem.2008.07.021.
The use of embryonic stem cell (ESC) differentiation to generate functional hepatic or pancreatic progenitors and as a tool for developmental biology is limited by an inability to isolate in vitro equivalents of regionally specified anterior definitive endoderm (ADE). To address this, we devised a strategy using a fluorescent reporter gene under the transcriptional control of the anterior endoderm marker Hex alongside the definitive mesendoderm marker Cxcr4. Isolation of Hex(+)Cxcr4(+) differentiating ESCs yielded a population expressing ADE markers that both can be expanded and is competent to undergo differentiation toward liver and pancreatic fates. Hex reporter ESCs were also used to define conditions for ADE specification in serum-free adherent culture and revealed an unexpected role for FGF signaling in the generation of ADE. Our findings in defined monolayer differentiation suggest FGF signaling is an important regulator of early anterior mesendoderm differentiation rather than merely a mediator of morphogenetic movement.
利用胚胎干细胞（ESC）分化来生成功能性肝或胰腺祖细胞，并将其作为发育生物学的工具，受到无法在体外分离出区域特异性前确定内胚层（ADE）等效物的限制。为了解决这个问题，我们设计了一种策略，使用在前肠内胚层标记物Hex以及确定中胚层标记物Cxcr4的转录控制下的荧光报告基因。分离Hex(+)Cxcr4(+)分化的ESC产生了一个表达ADE标记物的群体，该群体既可以扩增，又有能力向肝脏和胰腺命运分化。Hex报告ESC还用于确定无血清贴壁培养中ADE特化的条件，并揭示了FGF信号在ADE生成中的意外作用。我们在确定的单层分化中的发现表明，FGF信号是早期前中胚层分化的重要调节因子，而不仅仅是形态发生运动的介质。
