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影响大鼠胚胎多巴胺能神经元原代培养因素的研究 |
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陈立南 徐群渊 吕捷 张进禄
【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,PLL)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(1×105/cm2)分别种入由Matrigel及PLL作为基质的塑料培养皿中。体外培养5d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PLL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。
【关键词】 原代神经细胞培养；多巴胺能神经元；多聚左旋赖氨酸；免疫组织化学；Matrigel
【中图分类号】 Q421；Q8134.1+1 【文献标识码】 A 【文章编号】 0529-1356(2000)04-307
STUDY OF FACTORS AFFECTING PRIMARY CULTURE
OF DOPAMINERGIC NEURONS IN RAT FETUS
CHEN Li-nan
（Department of Anatomy,Beijing Medical University,Beijing 100083,China）
XU Qun-yuan Lü Jie ZHANG Jin-lu
（Beijing Institute for Neuroscience,Capital University of Medical Sciences,Beijing 100054,China）
【Abstract】 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 (1×105/cm2).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.
【Key words】 Primary neural cell culture; Dopaminergic neuron; Poly-L-lysine; Immunocytochemistry; Matrigel
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