摘要
为观察碱性成纤维细胞生长因子对体外培养的神经前体细胞的增殖和分化作用 ,本实验取胚胎 18d大鼠海马神经细胞 ,加入 2 5 ng/ m l碱性成纤维细胞生长因子置无血清培养基中进行培养。于培养第 4d和第 8d,用四唑盐比色法测定细胞活性 ,并用免疫组化方法定性、定量分析神经前体细胞的分裂和分化为神经元及神经胶质细胞的状态。结果显示 ,培养第 4d和第 8d,实验组的 OD值均增高 ,分别是对照组的 1.5倍和 1.8倍。免疫组化细胞分类计数显示 :培养第 4d,实验组神经前体细胞、神经元和少突胶质细胞数均明显增加 ,约是对照组的 2倍 ;但星形胶质细胞数无明显的变化。培养第 8d,实验组四类细胞均增加 ,约是对照组的 1.7倍。本实验提示 ,碱性成纤维细胞生长因子既能促进 E18的神经前体细胞的存活和分裂又能促进其向神经元和神经胶质细胞分化。并提示如拟在体外获得较大量和较高纯度的神经前体细胞 ,E18不是最理想的胚龄 ,需考虑选取胎龄更小的脑组织和加进足量的碱性成纤维细胞生长因子并进行传代培养。
In order to investigate effect of basic fibroblast growth factor(bFGF) on the prolifiration and differentiation of the neural progenitor of embryonic hippocampus of rats in vitro, 25 ng/ml bFGF was employed into the serum free medium of culture of hippocampal neural cells of embryonic day 18 rats in the present study. The effect of bFGF on the viability of cells cultured was detected by MTT colorimetric method and the effects of bFGF on proliferation and differentiation of hippocampal neural progenitors were analyzed qualitatively and quantitatively by means of immunochemistry, for the nestin, neurofilament, galactocerebroside and glial acidic fibrillary protein. The results showed that the OD value of experimental group was higher than that of control group by 1.5 and 1.8 times at 4 d and 8 d respectively. The quantitative analysis of each kinds of cells indicated that the number of neural progenitors, neurons and oligodendrocytes in experimental group were increased about 2 times as many as that in control group, but no differences of astrocytes between the two groups at 4 d. However, the number of four kinds of cells augmented about 1.7 times at 8 d. The results of this study suggest that bFGF can not only promote the survival, proliferation, but also facilitate the differentiation of neural progenitor of hippocampus to neurons and glial cells. To obtain more many purified neural progenitors in vitro, the embryonic day 18 is not an appropriate age. It is better to get younger embryo brain to culture and to add enough bFGF.
出处
《神经解剖学杂志》
CSCD
北大核心
2000年第3期201-204,共4页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金!( No. 3 9770 811
3 9670 775 )
国家教委跨世纪人才基金!(国教 95 -0 0 4)
广东省五个一兴医工程基金!(粤卫 1
关键词
BFGF
神经前体细胞
海马
细胞增殖
细胞分化
basic fibroblast growth factor
neural progenitor
culture
hippocampus
rat
