大鼠胚胎中脑多巴胺神经元的分离培养与鉴定

背景：神经细胞移植治疗是最有希望治愈帕金森病的方法之一。 目的：探讨大鼠胚胎中脑多巴胺神经元分离、培养与鉴定的方法。 方法：解剖分离E14d SD大鼠胚胎中脑组织,制备单细胞悬液,以神经细胞培养液培养,观察其生长情况并进行RT-PCR和免疫组织化学鉴定。 结果与结论：在神经细胞培养液中,细胞生长良好。RT-PCR和免疫组织化学结果显示大多数细胞表达多巴胺神经元特异性分子标志。证实实验成功建立了大鼠胚胎中脑多巴胺神经元分离培养与鉴定的方法。

Isolation,culture and identification of rat embryonic mesencephalic dopaminergic neurons

BACKGROUND:Nerve cell transplantation therapy is one of the most promising methods to cure Parkinson’s disease. OBJECTIVE:To investigate the culture, isolation and identification of rat embryo mesencephalic dopaminergic neurons. METHODS:The mesencephalon was dissected from rat embryo at embryonic day 14 (E14). The brain tissue was triturated into a fine single-cell suspension. The cells were cultured with the medium containing Neurobasal, B27 supplement, β-mercaptoethanol and fetal bovine serum. The cells were identified with RT-PCR as well as immunocytochemistry, respectively. RESULTS AND CONCLUSION:The cells survived well in the medium. RT-PCR and immunocytochemistry results showed mostly all neurons expressed molecular markers for dopaminergic neurons. The dopaminergic neurons derived from rat embryonic mesencephalon were successfully isolated and cultured, which will provide the foundation for the further research about cell therapy of Parkinson’s disease.