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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。
Objective To observe the effects of basic fibroblast growth factor, leukemia inhibitory factor, brain-derived neurotrophic factor and different combinations on the differentiation of neural stem cells from adult SD rats into neurons in vitro. Methods Adult SD rat brain neural stem cells were cultured in vitro with serum-free cell culture medium containing basic fibroblast growth factor (bFGF) and B27. Nestin immunocytochemical staining was performed on single-cell clones. According to the addition of nutritional factors The cells were divided into 5 groups: bFGF, LIF, BDNF, bFGF + LIF, bFGF + BDNF group. The 5 groups of cells were cultured for 1 week, NSE immunocytochemical staining was performed, and the ratio of positive cells was counted Statistical analysis. Results Compared with bFGF group, LIF group and BDNF group, the number of NSCs differentiated into neurons in bFGF + BDNF group and bFGF + LIF group was significantly higher than that in bFGF group (P <0.01) The proportion of nerve cells in bFGF + BDNF group was the highest. Conclusion Under the condition of bFGF culture, BDNF promotes the differentiation of neural stem cells into neurons in adult SD rats more than LIF.