小鼠胚胎干细胞在单层粘附培养中向神经细胞的分化

Differentiation of Mouse Embryonic Stem Cells into Neural Cells in Adherent Monoculture

目的 :探讨小鼠胚胎干 (ES)细胞在无血清培养基中以单层粘附培养方式向神经分化的方法。方法 :比较ES细胞在不同培养基中的生长情况 ,分析ES细胞在不同时间分化形成神经细胞的比例。结果 :( 1 )DMEM F1 2和Neurobasal B2 7的 1∶1混合培养基最适合ES的生长。 ( 2 )单层粘附的ES细胞表达神经细胞粘附分子 (NCAM)的比例随时间增长而增加 ,而nestin的表达先增加后下降。 ( 3)ES细胞可在两周分化为神经胶质及神经元 ,形成神经网络。结论 :小鼠ES细胞可在单层粘附培养中获得向神经的高效分化。

Objective : To evaluate different methods of adherent monocultured mouse embryonic stem (ES) cells' differentiation into neural cells in serum-free medium. Methods: We selected the best medium fitted for ES cells culture,and analyzed the neural differentiation of ES cells by comparing increased number of viable cells among different serum-free medium.Results: (1) DMEM/F12/Neurobasal/B27 was the optimal serum-free medium suitable for ES cells monoculture.(2) The percentage of cells positive for neural cell adhesion molecule (NCAM) increased along with the time,while the percentage of nestin positive cells began to increase at day 4 and then dropped at day 8.(3) Two weeks later,63 6%±7 8% of cultured cells expressed glial fibrillary acidic protein(GFAP),44 7%±6 8% cells expressed neurofilament (NF) and 35 7%±6 9% cells expressed Ⅲβ-tubulin.Conclusion: In adherent serum-free monoculture,ES cells can sufficiently develop into neural precursors and then differentiate into astrocytes and neurons.