不同细胞因子体外诱导神经干细胞向神经细胞分化的研究

目的观察碱性纤维母细胞生长因子、白血病抑制因子、脑源性神经营养因子及不同组合对成年SD大鼠脑神经干细胞在体外分化为神经细胞的作用。方法用含碱性纤维母细胞生长因子（bFGF）、B27的无血清细胞培养技术体外培养成年SD大鼠脑神经干细胞，单细胞克隆后行Nestin免疫细胞化学染色；根据培养液中所加营养因子的不同将单细胞克隆传代细胞分为5组培养：bFGF、LIF、BDNF、bFGF＋LIF、bFGF＋BDNF组，此5组细胞培养1周，进行NSE免疫细胞化学染色，计数阳性细胞比例后进行统计学分析。结果单细胞克隆培养后克隆球细胞表达Nestin；与bFGF组、LIF组、BDNF组相比，bFGF＋BDNF组和bFGF＋LIF组神经干细胞分化为神经细胞的比例较高（P〈0．01），其中bFGF＋BDNF组神经细胞的比例最高。结论在bFGF培养条件下，BDNF促进成年SD大鼠脑神经干细胞向神经细胞分化的能力高于LIF。

Inducement of rat neural stem cells to neuron in vitro with cytokines

Objective To evaluate the effects of bFGF, LIF, BDNF, and the differences from their groupings on neuronal differentiation of neural stem cells （NSCs） from CNS of adult rats in vitro. Methods The serum-free medium containing bFGF,B27 was used to culture NSCs isolated from CNS of adult rats. Immunocytochemistry for Nestin was used to identify monoclonal cells. The passaged monoclonal NSCs were divided into five groups, bFGF, LIF, BDNF, bFGF ＋ LIF and bFGF ＋ BDNF group. Immunocytochemistry for NSE was performed a week later to count the positive cells. Results The cultured monoclonalcells expressed nestin. The percentage of NSE positive cells differentiated from NSCs in bFGF ＋ LIF and EGF ＋ BDNF groups were much higher than those in other groups with the highest in bFGF ＋ BDNF group. Conclusion The ability of BDNF on promoting differentiation of NSCs from CNS of adult rats into neurons is higher than LIF in the media containing bFGF.