大鼠胚胎神经干细胞体外培养及诱导分化

Culture and Differentiation of Rat Embryonic Neural Stem Cells in Vitro

神经干细胞在理论研究和临床应用上有着广泛的前景.本文主要在体外分离培养SD大鼠胚胎前脑的神经干细胞,并分别用去除生长因子或添加全反式维甲酸(ATRA)两种方法诱导分化.免疫荧光染色技术分别检测细胞巢蛋白(Nestin)的表达及分化后β微管蛋白Ⅲ(β-Ⅲtubulin)、胶质纤维酸性蛋白(GFAP)的表达,计算分化率.结果显示:细胞生长状态良好,呈Nestin表达阳性.分化后可获得β-Ⅲtubulin及GFAP表达阳性的细胞,其中ATRA诱导方法获得β-Ⅲtubulin阳性细胞较多.

Neural stem cells（NSCs）have great potential in basic research and clinical application. In this study,NSCs were isolated from the forebrain of SD rat embryos,cultured in vitro,and then induced to differentiate by removing the growing factors or adding all-trans retinoic acid（ATRA）. Immunofluorescence technology was used to test the expression of Nestin and differentiation markerβ-Ⅲtubulin and GFAP,and then the percentage of differentiated cells were calculated. The results show that the isolated cells can be amplified in vitro and exhibit Nestin positive expression. The differentiated cells expressedβ-Ⅲtubulin and GFAP. Compared with growing factor withdrawal,addition of ATRA resulted in moreβ-Ⅲtubulin positive cells.