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

小鼠胚胎干细胞来源的神经嵴细胞的产生和分化分析

基本信息

批准号：
18591952
负责人：
MOTOHASHI Tsutomu
金额：
$ 2.53万
依托单位：
Gifu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591952/
关键词：
Neural Crest Cell Embryonic Stem Cell Sox10 Green Fluorescence Protein (GFP)Cell Sorter Neurocristopathy

项目摘要

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.

本研究的目的是利用小鼠胚胎干细胞衍生的神经嵴细胞，分析神经嵴细胞的分化和移动性。为了纯化胚胎干细胞衍生的神经嵴样细胞，我们建立了SoxlO-ires-gfp胚胎干细胞。SoxlO-ires-gfp ES细胞在sox10基因停止密码子后含有绿色荧光蛋白（GFP）基因。当胚胎干细胞分化为神经嵴细胞时，细胞与体内神经嵴细胞中表达的Sox10一起表达GFP，利用GFP表达纯化胚胎干细胞衍生的神经嵴细胞。利用SoxlO-ires-gfp胚胎干细胞，我们从培养中有效富集胚胎干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移动性。我们分析了胚胎干细胞中神经嵴样细胞的发育情况。结果表明，神经冠样细胞可分为Sox10+/c-Kit-和Sox10+/c-Kit+群体，其中Sox10+/c-Kit-群体比Sox10+/c-Kit+群体更不成熟。此外，我们建立了来自SoxlO-ires-gfp ES细胞的小鼠。在胚胎中，神经嵴细胞及其衍生物被绿色荧光蛋白清晰地标记。SoxlO-ires-gfp ES细胞和来源于SoxlO-ires-gfp ES细胞的小鼠被认为是神经嵴研究的有价值的工具。神经嵴细胞。