人胚神经干细胞的分离、培养与鉴定  

Isolation, Cultivation and Identification of Neural Stem Cell from Human Embryonic CNS

作　　者：王兰[1,2] 胡火珍[1] 张承虎[3] 李小玉[1] 陶大昌[1] 陈芬[1] 

机构地区：[1]四川大学华西医学中心细胞生物学教研室 [2]泸州医学院生物教研室,646000 [3]泸州医学院计算机管理科

出　　处：《生物医学工程学杂志》2002年第2期264-267,共4页Journal of Biomedical Engineering

摘　　要：为了探讨人胚神经干细胞体外培养条件和分化情况 ,摸索出一种切实可行的获得较纯、多潜能人胚神经干细胞的方法。我们取三月龄人胎脑 ,用胰蛋白酶消化法分离单个细胞 ,部分冻存 ,另一部分进行细胞培养 ,加EGF、b FGF刺激生长 ,有限稀释法获得单细胞克隆 ,血清诱导分化 ,并用免疫组化方法进行鉴定。结果显示 ,EGF和 b FGF同时存在的无血清培养基中有大量神经干细胞团生成 ,含血清培养基则诱导神经干细胞分化成为神经元、星型胶质细胞、少突胶质细胞。这表明 ,神经干细胞的存活和分裂有赖于 EGF和 b FGF的共同作用。This is a study on the cultivation condition in vitro and differentiation of neural stem cells from human embryonic brain in order to find a way to get purified multipotential neural stem cells. The single cells was derived from the three month embryonic brain digested with trypsin ,some cells was frozen, the other cells were expanded with EGF and bFGF, the single cell clone was obtained by the way of limited dilution, and the serum was used to induce the cells differentiation. The cells were detected with the method of immunohistochemistry. The results showed that a lot of neurospheres could be seen in the presence of mitogens (both EGF and bFGF) and serum could induce neural stem cells to differentiate into neurons, astrocytes, and oligodendrocytes. These indicate that the survival and proliferation of neural stem cells rely on the cooperation of EGF and bFGF. The neural stem cells can also be harvested from the frozen cells.

关 键 词：人胚 神经干细胞 细胞培养 细胞冻存 BFGF EGF 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]