摘要
采用类胚体法(EB)的EB分化4-/4+程序和单层分化法研究不同的实验方法对胚胎干细胞神经分化的影响。用4-/4+分化程序进行胚胎干细胞的神经分化时,饲养层细胞的存在对类胚体的形成无影响,在神经分化的13d,仍可观察到46C细胞的Sox1-GFP绿色荧光,此时的免疫染色结果显示,分化细胞中除了有Nestin阳性的神经干细胞及胶质纤维酸性蛋白(GFAP)阳性的神经胶质细胞外,以β-tubulin 阳性神经元细胞最为多见,这些神经元呈现出多种形态;用单层分化法进行胚胎干细胞(ES)细胞的神经分化至13d时,分化形成的神经细胞种类与类胚体法相同,但神经元的细长纤维形成了复杂的网状结构,提示两种分化方法获得的神经元处于不同发育阶段。研究结果表明,两种分化方法均能为ES细胞提供从神经分化的起始到继续分化所需的环境,EB分化法适合于大量地获取分化的细胞,而单层分化法可直观地研究分化过程的细胞变化。
The neural differentiation process of mouse embryonic stem (ES) cell induced by embryoid differentiation and monolayer differentiation was investigated in this study. When embryoid differentiation method was employed by 4-/4+retinoic acid(RA) treatment and trypsinization of cellular aggregates before culture in dulbecco's modified eagle medium(DMEM)/F12 plus N2/B27 medium,Sox1-GFP still expressed at 13 d. Nestin positive cells were more than glial fibrillary acid protein(GFAP) positive.β-tubulin Ⅲ positive cells were still dominated which gave multiplex morphology. When monolayer differentiation method was employed by culturing embryonic stem(ES) cell in dulbecco's modified eagle medium(DMEM)/F12 plus N2/B27 medium, at 13 d many complex axonal networks formed.Among differentiated neural cells,β-tubulin Ⅲ positive were predominate and some were nestin positive and GFAP positive. The results indicate that both methods can be employed to induce the neural differentiation of mouse ES cells effectively.
出处
《广西农业生物科学》
CAS
CSCD
2004年第4期329-332,共4页
Journal of Guangxi Agricultural and Biological Science
基金
国家863计划资助项目(2002AA206605)
第21期广西出国留学基金资助项目
关键词
胚胎干细胞
神经分化
类胚体
单层分化法
embryonic stem cell
neural differentiation
embryoid body
monolayer differentiation
