Analysis of generation and differentiation of neural crest cells derived from mouse embryonic stem cells

本研究的目的是利用小鼠胚胎干细胞来源的神经嵴细胞，分析神经嵴细胞的分化和迁移能力。为了纯化ES细胞来源的神经嵴样细胞，我们建立了SoxlO-ires-gfp ES细胞。Sox 10-ires-gfp ES细胞在sox 10基因的终止密码子后具有绿色荧光蛋白（GFP）基因。当ES细胞分化为神经嵴细胞时，该细胞表达GFP以及在体内神经嵴细胞中表达的Sox 10，并且通过利用GFP表达纯化ES细胞来源的神经嵴细胞。使用SoxlO-ires-gfp ES细胞，我们有效地从培养物中富集了ES细胞衍生的神经嵴样细胞。将富集的神经嵴样细胞移植到鸡胚中，表明神经嵴样细胞具有与体内神经嵴细胞相似的运动性。并对ES细胞向神经嵴样细胞的分化进行了分析。结果表明，Sox 10 +/c-Kit-和Sox 10 +/c-Kit+两种细胞群中，Sox 10 +/c-Kit-细胞群比Sox 10 +/c-Kit+细胞群具有更多的未成熟神经嵴样细胞。在胚胎中，神经嵴细胞及其衍生物被绿色荧光蛋白清楚地标记。SoxlO-ires-gfp ES细胞和衍生自SoxlO-ires-gfp ES细胞的小鼠被认为是用于神经嵴研究的有价值的工具。神经嵴细胞。

The purpose of this research is to analyze the differentiation and mobility of neural crest cells by utilizing the mouse ES cells derived neural crest cell. We established SoxlO-ires-gfp ES cells in order to purify the ES cell derived neural crest like cells. The SoxlO-ires-gfp ES cells have the gene of green fluorescence protein (GFP) after the stop codon of sox10 gene. When the ES cells are differentiated into neural crest cells, the cells express GFP together with Sox10, which is expressed in in vivo neural crest cells, and the ES cell derived neural crest cells are purified by utilizing GFP expression. Using the SoxlO-ires-gfp ES cells, we effectively enriched the ES cell derived neural crest-like cells from culture. Transplantation of the enriched neural crest-like cells into chick embryo suggested that the neural crest-like cells had similar mobility to in vivo neural crest cells. And we analyzed the development of the neural crest-like cells from ES cells. We showed that the neural crest-like cells were categorized into the Sox10+/c-Kit- and the Sox10+/c-Kit+ population, and the Sox10+/c-Kit- population especially was more immature neural crest-like cells compare to the Sox10+/c-Kit+ population.Furthermore, we established the mice derived from the SoxlO-ires-gfp ES cells. In the embryos, neural crest cells and the derivatives were clearly marked with Green fluorescence protein. The SoxlO-ires-gfp ES cells and the mice derived from the SoxlO-ires-gfp ES cells are thought to be valuable tools for neural crest studies. Neural Crest Cell.