小胎龄鼠的腹侧中脑神经前体细胞体外培养研究

目的体外培养小胎龄的胎鼠腹侧中脑神经前体细胞,根据多巴胺能神经元产量,确定较好的研究对象,为细胞移植治疗帕金森病奠定基础。方法取不同胎龄胎鼠(E10、E11、E12)的腹侧中脑神经前体细胞,体外加碱性成纤维细胞生长因子扩增7d后用含AA-2P的分化增养液培养7 d,行免疫荧光染色比较多巴胺能神经元形成情况。结果E10、E11、E12细胞经培养后细胞总数分别增加76.39倍、59.67倍、51.85倍。多巴胺能神经元占所有细胞的比例在E11、E10、E12依次显著降低。对于多巴胺能神经元占所有神经元的比例,E11显著高于E10和E12。结论来自胎龄11 d的胎鼠腹侧中脑神经前体细胞是较好的体外培养对象,可为细胞移植研究提供较为充足的细胞资源。

Study on in vitro culture of ventral mesencephalic precursors from rat embryos at young embryonic stages

Objective Ventral mesencephalic dopaminergic precursors from rat embryos at young embryonic stages were cultured in vitro. The yields of dopaminergic neurons (DNs) determined the optimal research material for the subsequent study on cell transplantation as a treatment of Parkinson's disease. Methods Ventral mesencephalic precursors from rat embryos at different embryonic stages (from E10, E11, E12) proliferated with bFGF for 7 days and differentiated for the next 7 days in the special differentiation medium containing L-ascorbic acid-2-phosphate sesquimagnesium salt (AA-2P). The yields of DNs were compared by means of immunofluorescence staining. Results The total cell numbers in E10, E11 and E12 cultures increased about 76.39 times, 59.67 times and 51.85 times, respectively. The ratio of DNs versus the total cell population in E11, E10 and E12 cultures decreased sequentially and significantly. The ratio of DNs versus all the neurons in E11 culture was significantly higher than those in E10 and E12 cultures. Conclusion Ventral mesencephalic precursors from E11 rat embryos are the optimal research material, which can in vitro supply enormous cell resources for the study of cell transplantation.