近交系大鼠胚胎中脑前体细胞体外培养初步研究

Preliminary study on the culture of ventral mesencephalic precursors derived from inbred rat embryos

目的 尝试体外培养近交系大鼠胚胎中脑前体细胞（ventral meseneephalic precursor，VMP）并进行诱导分化。方法 取材E11近交系大鼠胚胎腹侧中脑细胞在bFGF培养液中原代培养7天，撤去bFGF，加AA-2P诱导分化7～10天，免疫荧光染色法分析前体细胞分化情况。结果 体外培养14天及17天，细胞数量分别扩增至培养前的49．76倍和58．43倍，免疫荧光染色显示细胞群体中除了VMP外，还可见大量的神经元（包括多巴胺能神经元）和星形胶质细胞等。结论 近交系大鼠VMP能够在体外扩增并分化，可得到较多的多巴胺能神经元用于后续分子生物学研究。

Objective To culture ventral mesencephalic precursors （VMPs） derived irom inbred rat embryos in vitro. Methods VMPs harvested from ventral mesencephalons of E11 inbred rat embryos were cultured in the medium supplemented with bFGF for 7 days, and differentiated into dopaminergic neurons in free-bFGF medium supplemented with AA-2P for 7 - 10 days. Immunofluorescence staining was performed to evaluate the outcome of induced differentiation. Results The numbers of VMPs multiplied about 49. 76 and 58. 43 folds afer VMPs Were cultared for 14 and 17 days, respectively. Immunofluorescence staining showed that the cell popu- lation was comprised of not only VMPs but also many neurons and astrocytes differentiated from the VMPs. Conclusion VMPs of inbred rat embryos can proliferate and differentiate well in vitro,producing considerable dopaminergic neurons for subsequent research.