细胞来源和培养液成分对大鼠神经前体细胞增殖和分化的影响

Influence of origins of cells and compositions of the culture medium on proliferation and differentiation of neural precursor cells of rat

在成功建立体外分离培养大鼠胚胎脑和脊髓神经前体细胞(neuron precursor cells,NPCs)的基础上,本研究设计了三种培养液组合:DF/N2、DF/B27和DF/(N2+B27),观察在不同培养液成分对胚胎脑和脊髓NPCs增殖和分化的影响。结果显示:与NF/N2組和DF/B27组相比,脑来源的NPCs在DF/(N2+B27)中增殖最快、最稳定(P<0.01),而脊髓来源的NPCs在三种培养液组合中的增殖速度无明显差异。脑和胚胎15 d脊髓来源的NPCs在DF/B27和DF/(N2+B27)中分化为神经元的比例明显高于DF/N2组合(P<0.01);取自胚胎15 d的脊髓NPCs分化为神经元和少突胶质细胞的比例均显著高于胚胎16 d的NPCs(P<0.05)。以上结果提示:(1)在培养液中同时添加N2和B27不仅可以提高体外培养的NPCs的增殖速度,同时可显著增加神经元分化的比例;(2)NPCs的分化潜能可因NPCs来源(脑或脊髓)和发育阶段的不同而有差异。

We have establish the culture system of isolation and cultivation of the neural precursor cells （NPCs） from the embryonic rat brain and spinal cord. Here we used three medium combinations ： DF/N2, DF/B27 and DF/（ N2 ＋ B27 ）, to observe the influence of each medium combination on the proliferation and differentiation of NPCs from embryonic rat brain and spinal cord. The result are as follows： The NPCs, from embryonic rat brain, cultured in DF/（ N2 ＋ B27 ） showed high efficient expansion and more stability than that in DF/N2 and DF/B27 groups （ P 〈 0.01 ）. However, the proliferations of NPCs from embryonic rat spinal cord, in the three medium combinations, were no difference. The percents of neurons differentiated from NPCs of brain and E15 spinal cord were significantly higher in DF/B27 and DF/（ N2 ＋ B27 ） groups compared with DF/N2 group （ P 〈 0.01 ） ; The percents of neurons and oligodendroeytes differentiated from NPCs of E15 spinal cord were higher than that of E16 spinal cord （ P 〈 0.05 ）. All these results indicate that ： （ 1 ） Adding N2 and B27 into culture medium could not only promote the proliferation rapid of NPCs from embryonic rats, but could also increase the percents of neurons differentiated from NPCs ; （ 2 ） The NPCs from different part of CNS （ brain or spinal cord） or different developing stage, would have different differentiation potential.