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Study on cell lineage of neuronal stem cell in vitro

神经干细胞体外细胞谱系研究

基本信息

批准号：
14580778
负责人：
YAMANOHA Banri
金额：
$ 2.24万
依托单位：
Soka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580778/
关键词：
Neural stem cells Neural precursor cell Flow cytometry Primary culture basic FGF Neurospheres EGF Differentiation 前駆細胞 Neurospheres bFGF

项目摘要

Basic fibroblast growth factor(bFGF) and epidermal growth factor(EGF) stimulates the proliferation of neural stem cells from early developmental brain. For the receptor of the growth factor on the cells, the timing of expression in culture system is seem to affects its cell lineage. In this study, focus on cell lineage of neural stem cells. The cells are separated from forebrain in embryonic stage at 12 day. The cell isolation method is established to use flow cytometry with light scattering. Isolation qualify in this method would be highly to purify neural stem cells from the cell suspension which effects to cellular events in culture system. In cell suspension of forebrain, A2B5 bound cells were appeared 12% in total cells, whereas almost cells were belonged to nestin-positive cells.The bound cells in both antibody were also appeared in the suspension. Cell culture is performed in differentiation medium in cells isolated by flow cytometry, respectively. These cell populations were generate neuronal cells first and glial cells later. The cells generate neural progenitor and glial progenitor cells randomly. This result would seem that glial progenitor was not accepted FGF signaling in culture period at 8 to 10 days.In medium containing EGF, neural stem cells were appeared neurospheres at 8 to 10 days, whereas not before 8 day. It seem to be a different between FGF and EGF stimulate cells.

碱性成纤维细胞生长因子（bFGF）和表皮生长因子（EGF）刺激早期发育的大脑神经干细胞的增殖。对于细胞上的生长因子受体，在培养系统中的表达时机似乎影响其细胞系。本研究的重点是神经干细胞的细胞谱系。胚胎期第12天将细胞与前脑分离。建立了光散射流式细胞术分离细胞的方法。该方法能有效地从细胞悬浮液中纯化神经干细胞，并对培养系统中的细胞事件产生影响。在前脑细胞悬液中，A2B5结合细胞占总细胞的12%，而几乎所有细胞都属于巢蛋白阳性细胞。两种抗体的结合细胞也出现在悬浮液中。流式细胞术分离的细胞分别在分化培养基中进行细胞培养。这些细胞群首先产生神经细胞，然后产生神经胶质细胞。细胞随机生成神经祖细胞和神经胶质祖细胞。这一结果表明，在培养8 ~ 10 d时，胶质祖细胞未接受FGF信号。在含有EGF的培养基中，神经干细胞在8 ~ 10天出现神经球，而在8天之前没有出现。FGF和EGF刺激细胞似乎是不同的。