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Modeling early stages of endoderm development in epiblast stem cell aggregates with supply of extracellular matrices.
Development, Growth & Differentiation ( IF 1.7 ) Pub Date : 2020-05-08 , DOI: 10.1111/dgd.12663 Sachiko Inamori 1 , Mai Fujii 1 , Sayaka Satake 1 , Hideaki Iida 1 , Machiko Teramoto 1 , Tomoyuki Sumi 2 , Chikara Meno 2 , Yasuo Ishii 1, 3 , Hisato Kondoh 1
Development, Growth & Differentiation ( IF 1.7 ) Pub Date : 2020-05-08 , DOI: 10.1111/dgd.12663 Sachiko Inamori 1 , Mai Fujii 1 , Sayaka Satake 1 , Hideaki Iida 1 , Machiko Teramoto 1 , Tomoyuki Sumi 2 , Chikara Meno 2 , Yasuo Ishii 1, 3 , Hisato Kondoh 1
Affiliation
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Endoderm precursors expressing FoxA2 and Sox17 develop from the epiblast through the gastrulation process. In this study, we developed an experimental system to model the endoderm-generating gastrulation process using epiblast stem cells (EpiSCs). To this end, we established an EpiSC line i22, in which enhanced green fluorescent protein is coexpressed with Foxa2. Culturing i22 EpiSCs as aggregates for a few days was sufficient to initiate Foxa2 expression, and further culturing of the aggregates in Matrigel promoted the sequential activation of transcription factor genes involved in endoderm precursor development, e.g., Eomes, Gsc, and Sox17. In aggregation culture of i22 cells for 3 days, all cells expressed POU5F1, SOX2, and E-cadherin, a signature of the epiblast, whereas expression of GATA4 and SOX17 was also activated moderately in dispersed cells, suggesting priming of these cells to endodermal development. Embedding the aggregates in Matrigel for further 3 days elicited migration of the cells into the lumen of laminin-rich matrices covering the aggregates, in which FOXA2 and SOX17 were expressed at a high level with the concomitant loss of E-cadherin, indicating the migratory phase of endodermal precursors. Prolonged culturing of the aggregates generated three segregating cell populations found in post-gastrulation stage embryos: (1) definitive endoderm co-expressing high SOX17, GATA4, and E-cadherin, (2) mesodermal cells expressing a low level of GATA4 and lacking E-cadherin, and (3) primed epiblast cells expressing POU5F1, SOX2 without E-cadherin. Thus, aggregation of EpiSCs followed by embedding of aggregates in the laminin-rich matrix models the gastrulation-dependent endoderm precursor development.
中文翻译：
模拟外胚层干细胞聚集体中内胚层发育的早期阶段，并提供细胞外基质。
表达FoxA2和Sox17的内胚层前体通过上皮过程从上皮细胞发育而来。在这项研究中，我们开发了一个实验系统，用于使用表皮干细胞（EpiSCs）来模拟产生内胚层的胃形成过程。为此，我们建立了EpiSC系i22，其中增强的绿色荧光蛋白与Foxa2共表达。将i22 EpiSC作为聚集体培养几天就足以启动Foxa2表达，并且进一步在Matrigel中培养聚集体可促进涉及内胚层前体发育的转录因子基因（例如Eomes，Gsc和Sox17）的顺序激活。在i22细胞聚集培养3天后，所有细胞均表达POU5F1，SOX2和E-钙黏着蛋白（表皮细胞的标志），而GATA4和SOX17的表达在分散的细胞中也被适度激活，表明这些细胞向内胚层发育致敏。将聚集体在Matrigel中再包埋3天，引起细胞迁移到覆盖聚集体的富含层粘连蛋白的基质腔中，其中FOXA2和SOX17以高水平表达，同时伴随着E-钙粘蛋白的丢失，表明迁移阶段内胚层前体。长时间的聚集体培养产生了三个在分离后胚胎中发现的分离细胞群：（1）共表达高SOX17，GATA4和E-钙黏着蛋白的定形内胚层，（2）表达低水平且缺乏E的中胚层细胞-钙黏着蛋白，和（3）引发表达POU5F1，SOX2而没有E-钙黏着蛋白的上皮细胞。从而，
更新日期：2020-04-11
中文翻译：
模拟外胚层干细胞聚集体中内胚层发育的早期阶段，并提供细胞外基质。
表达FoxA2和Sox17的内胚层前体通过上皮过程从上皮细胞发育而来。在这项研究中，我们开发了一个实验系统，用于使用表皮干细胞（EpiSCs）来模拟产生内胚层的胃形成过程。为此，我们建立了EpiSC系i22，其中增强的绿色荧光蛋白与Foxa2共表达。将i22 EpiSC作为聚集体培养几天就足以启动Foxa2表达，并且进一步在Matrigel中培养聚集体可促进涉及内胚层前体发育的转录因子基因（例如Eomes，Gsc和Sox17）的顺序激活。在i22细胞聚集培养3天后，所有细胞均表达POU5F1，SOX2和E-钙黏着蛋白（表皮细胞的标志），而GATA4和SOX17的表达在分散的细胞中也被适度激活，表明这些细胞向内胚层发育致敏。将聚集体在Matrigel中再包埋3天，引起细胞迁移到覆盖聚集体的富含层粘连蛋白的基质腔中，其中FOXA2和SOX17以高水平表达，同时伴随着E-钙粘蛋白的丢失，表明迁移阶段内胚层前体。长时间的聚集体培养产生了三个在分离后胚胎中发现的分离细胞群：（1）共表达高SOX17，GATA4和E-钙黏着蛋白的定形内胚层，（2）表达低水平且缺乏E的中胚层细胞-钙黏着蛋白，和（3）引发表达POU5F1，SOX2而没有E-钙黏着蛋白的上皮细胞。从而，
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