EGF和bFGF对成年大鼠神经干细胞增殖和分化的影响

本研究旨在探索可促进成年大鼠神经干细胞增殖并形成较多的克隆球以及其分化出较多神经元的因素。取成鼠前脑室下区的组织进行原代培养 ,将之分为三组分别加入 EGF、b FGF以及 EGF+ b FGF,观察克隆球的形成状况。一周后收集三组原代细胞克隆球 ,加入完全培养液 (仅含 10 %胎牛血清 )进行分化实验。分化 14 d后 ,分别用 MAP-2和 GFAP的单克隆抗体进行免疫荧光标记 ,计算阳性细胞数量。无血清培养结果显示 ,b FGF组和 EGF+ b FGF组原代培养液中形成的原代克隆球数量和直径的差别不明显 ,但都明显地大于 EGF组。免疫荧光结果显示 ,b FGF组和 EGF+ b FGF组中的克隆球分化出 MAP-2阳性神经元的数量明显多于 EGF组 ,而 EGF组则能产生较多的胶质细胞。提示 ,b F GF能促进成年大鼠神经干细胞增殖 ,所形成的细胞克隆球能分化为较多的神经元。

THE EFFECT OF EGF AND bFGF ON THE PROLIFERATION AND DIFFERENTIATION OF NEURAL STEM CELLS FROM ADULT RATS

To seek the factors which can achieve more neural stem cell clones from adult rats and produce more neurons, the neural stem cells isolated from subventricular zone (SVZ) of adult rats were cultured in the medium containing EGF, bFGF and both. After 7 days, these neurospheres were collected respectively and differentiated in the culture medium only containing 10% fetal bovine serum. After 14 days, the differentiated cell clones were detected immunocytochemically using antibodies of specific marker proteins, MAP 2 for neurons and GFAP for astrocytes. The serum free culture results indicated that the number and diameter of the primary clones showed no distinct difference between the bFGF and EGF+bFGF group. The number of the primary clones were more and the diameter were longer in the bFGF and EGF+bFGF groups than that in the EGF group. The immunofluorescence results showed that the clones in the bFGF and EGF+bFGF groups could differentiate more neurons than that in the EGF group. There were many astrocytes in the EGF group. Our study suggests that bFGF can promote the proliferation of neural stem cells from adult rats and these clones can differentiate into more neurons.