影响大鼠胚胎多巴胺能神经元原代培养因素的研究  

STUDY OF FACTORS AFFECTING PRIMARY CULTURE OF DOPAMINERGIC NEURONS IN RAT FETUS

作　　者：陈立南[1] 徐群渊[2] 吕捷 张进禄[2] 

机构地区：[1]北京医科大学解剖学系,北京100083 [2]首都医科大学北京神经科学研究所,北京100054

出　　处：《解剖学报》2000年第4期307-312,I006,共7页Acta Anatomica Sinica

摘　　要：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 。Objective\ In order to obtain high purity of primary culture of dopaminergic neurons in rat fetus,we tried to modify the culture procedure.Using improved techniques,we compared the growth status of dopaminergic neurons cultured on Matrigel and PLL(Poly\|L\|Lysine)substrates. Methods\ The DA neurons were obtained from the ventral mesencephalon of SD rat embryos(E13) by using a different dissecting method and then plated in plastic dishes coated with Matrigel and PLL substrates in high density (1×10\+5/cm\+2).After cultured for 5 days in vitro, TH immunocytochemical staining was performed and length of TH immunoreactive neurites was measured. Results and Conclusions\ 1\^Cultured dopaminergic neurons in high\|purity(dopaminergic neuron/all neuron=15%) could be got by using this improved culture method.2\^ Matrigel substrate promoted neurites outgrowth of cultured neurons more potently than PLL substrate,but did not increase the survival of the neurons.\;

关 键 词：原代神经细胞培养 多巴胺能神经元 PLL MATRIGEL 

分 类 号：R338[医药卫生—人体生理学] R329－33[医药卫生—基础医学]