胎鼠中脑多巴胺细胞混合培养方法探索

目的 探索一种新的胚胎大鼠腹侧中脑黑质细胞的混合培养方法,以获得具有高比例多巴胺神经元的原代细胞体系,有利于在体外条件下进行帕金森病(Parkinson's disease,PD)的发病机制和防治的研究.方法 分离E15 (Embryonic Day 15)SD胎鼠中脑黑质区域组织,分散为单细胞后,分别用DMEM/F12+ 10％FBS含血清培养基和Neurobasal+ N1无血清培养基设置不同组别进行培养,通过免疫组织化学方法检测在不同培养条件下细胞的生长状态及多巴胺能神经元的比例.结果 DMEM/F12+ 10％ FBS与Neurobasal+ N1先后一周交替换液的的培养体系中获得的TH阳性神经元占神经元的比例可达31％左右,明显高于单独使用DMEM/F12+ 10％ FBS培养组(约10％).结论 DMEM/F12+ 10％ FBS与Neurobasal +N1先后一周交替换液是一种黑质神经元和胶质细胞混合培养并能获得高比例多巴胺神经元的有效培养方法.

Experimental study on the mixed culture of embryonic rat ventral mesencephalic dopaminergic cells

Objective To explore an effective method for embryonic rat ventral mesencephalic tissue culture to obtain high proportion of dopaminergic neurons which are feasible for in vitro studies on the pathogenesis and treatment of Parkinson＇s disease. Methods Ventral mesencephalic cells derived from El5 （Embryonic Day 15）SD fetal rats＂ substantia nigra. The cells were cultured for 2 weeks in DMEM/F12 q- 10}＇~ FBS and Neurobasal -~ N1 serum-free medium, respectively. The growth status was observed and the rate of dopaminergic neurons in different culture conditions was detected by immunohistochemistry. Results The rate of TH positive neurons in the group with DMEM/F122＋10%FBS followed with Neuro- basalq-N1 （over 30%） was much higher than that of the group only with DMEM/F12＋10% FBS（about 10%）. Conclusion Comparatively high proportion of dopamergic neurons from substantia nigra tissue can be obtained under the culture condition of DMEM/F12q-FBS followed with Neurobasal＋N1.