神经球方法体外无血清含生长因子培养的神经干细胞主要向神经胶质细胞分化

Differentiation of neural stem cells into glial cells cultured in serum-free medium containing growth factors by using neurosphere assay

背景:体外分离培养出神经干细胞并掌握其生物学特性是神经干细胞移植研究的前提。目的:采用神经球方法分离培养胎鼠皮质神经干细胞,并观察其生长和分化特性。方法:利用无血清悬浮方法分离培养胚胎来源的神经干细胞,用免疫细胞化学的方法检测神经干细胞及其分化细胞的标志物。结果与结论:体外无血清含生长因子的培养液培养胎鼠皮质神经干细胞,可得到悬浮生长的神经球,但悬浮生长的神经球可以发生融合,免疫细胞化学显示神经球呈巢蛋白表达阳性,神经干细胞可以分化为神经元,神经胶质细胞和少突胶质细胞,但主要向神经胶质细胞分化。说明在血清含有生长因子的而培养条件下,利用神经球方法可以分离培养出胎鼠皮质神经干细胞。

BACKGROUND： Isolation and culture of neural stem cells in vitro and to master their biological characteristics are the basis of neural stem cells transplantation. OBJECTIVE： To isolate and culture cerebral cortex neural stem cells from rat embryos by neurosphere assay, and to observe the growth and differentiation characteristics of neural stem cells. METHODS： Neural stem cells were isolated from rat embryonic cerebral cortex and cultured in serum-free medium. Immunocytochemical staining was used to examine neural stem cells and differentiated cells markers. RESULTS AND CONCLUSION： Neural stem cells grew as free-floating neurospheres in the serum-free conditions, but spontaneous fusion occurred among some neurospheres. Immunocytochemical staining showed these neruospheres positively expressed nestin, and neural stem cells could differentiated into neurons, astrocytes and oligondendrocytes, but mainly differentiated into astrocytes. In serum-free medium containing growth factors, cerebral cortex neural stem cells could be isolated and cultured by using neurosphere assay.