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目的 试图通过方法改进获得高纯度原代多巴胺能神经元 ,并研究 Matrigel及多聚左旋赖氨酸(poly- l- lysine,PL L )等不同基质对于大鼠胚胎 13d中脑黑质神经元突起生长的不同作用。 方法 无菌技术取得孕 13d SD大鼠胚胎 ,采用改进的取材方法 ,得到胎鼠腹侧中脑细胞 ,以较高密度 (1× 10 5 / cm2 )分别种入由 Matrigel及 PL L作为基质的塑料培养皿中。体外培养 5 d后 ,对培养物进行 TH免疫细胞化学染色 ,测量 TH免疫反应突起的长度并计算 TH免疫反应神经元占细胞总数的比例。 结果和结论 1.采用改进的方法 ,能够得到高纯度的多巴胺能神经元 (多巴胺神经元占培养细胞总数的比例为 15 % ) ;2 .与 PL L相比 ,Matrigel能有效地促进多巴胺能神经元突起的生长 ,但不能特异地促进中脑培养细胞的存活
OBJECTIVE: To improve the purity of primary dopaminergic neurons and to investigate the effects of Matrigel and PL-L on the neurite outgrowth The different effects of growth. Methods The fetal rat embryonic embryo was obtained by aseptic technique. The ventral mesencephalic cells of fetus were obtained by improved material extraction method. Matrigel and PL L were used as the matrix in higher density (1 × 10 5 / cm 2) Plastic Petri dishes. After cultured for 5 days in vitro, the cultures were subjected to TH immunocytochemical staining to measure the length of the TH immunoreactive protrusions and to calculate the ratio of TH immunoreactive neurons to the total number of cells. RESULTS AND CONCLUSIONS 1. High-purity dopaminergic neurons (dopamine neurons account for 15% of total cultured cells) can be obtained by an improved method; 2. Compared with PL L, Matrigel can effectively promote dopaminergic neurons Protuberant growth, but can not specifically promote the survival of cultured mesencephalon cells