Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells
Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells
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DOI：
10.1038/nbt1167
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发表时间：
2005-12-01
影响因子：
46.9
通讯作者：
Nishikawa, SI
中科院分区：
文献类型：
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作者：
Yasunaga, M;Tada, S;Nishikawa, SI
从胚胎干细胞（ES细胞）中高纯度制备特定谱系需要选择性培养条件以及用于引导和监测分化的标志物。在这项研究中，我们利用一种小鼠ES细胞系区分了定形内胚层和脏壁内胚层，该细胞系在 Goosecoid（Gsc）和Sox17基因座分别携带绿色荧光蛋白（gfp）和人白细胞介素2受体α（也称为CD25）标记基因。这种细胞系使我们能够监测Gsc(+)Sox17(+)定形内胚层和Gsc(-)Sox17(+)脏壁内胚层的产生，并确定能差异诱导定形内胚层和脏壁内胚层的培养条件。通过比较定形内胚层和脏壁内胚层的基因表达谱，我们鉴定出在这两种群体中差异表达的七种表面分子。七种标志物之一的Cxcr4有单克隆抗体可用，这使我们能够在选择定形内胚层的条件下，从未经基因操作的ES细胞中监测和纯化Gsc +群体。
Preparation of specific lineages at high purities from embryonic stem (ES) cells requires both selective culture conditions and markers to guide and monitor the differentiation. In this study, we distinguished definitive and visceral endoderm by using a mouse ES cell line that bears the gfp and human IL2R alpha (also known as CD25) marker genes in the goosecoid (Gsc) and Sox17 loci, respectively. This cell line allowed us to monitor the generation of Gsc(+)Sox17(+) definitive endoderm and Gsc(-)Sox17(+) visceral endoderm and to define culture conditions that differentially induce definitive and visceral endoderm. By comparing the gene expression profiles of definitive and visceral endoderm, we identified seven surface molecules that are expressed differentially in the two populations. One of the seven markers, Cxcr4, to which a monoclonal antibody is available allowed us to monitor and purify the Gsc+ population from genetically unmanipulated ES cells under the condition that selects definitive endoderm.