酶联合消化法培养小鼠主动脉平滑肌原代细胞

Primary culture of mouse aortic smooth muscle cells by enzyme-combined digestion

目的建立一种适用于体外有效分离培养小鼠主动脉血管平滑肌细胞的技术方法。方法分离主动脉,用优化的酶联合消化法获取主动脉平滑肌细胞,用形态学法、免疫细胞化学法鉴定。结果该方法所分离的主动脉平滑肌细胞生长旺盛,细胞活性好,7～9d后细胞总量可达到5×105个/瓶,细胞纯度可达85%以上,细胞呈长梭形、放射状、束状生长,平行排列呈峰谷状,免疫荧光显示胞浆内α-SMA蛋白表达阳性。结论该酶联合消化法可在体外短时间内获取数量可观、高纯度的主动脉平滑肌细胞。

Objective To establish a technical method which could effectively isolate and culture mouse aortic vascular smooth muscle cells in vitro.Methods The aortic smooth muscle cells were harvested by optimization enzyme-combined digestion with separated aorta.These cells were identified using morphological method and immunocytochemistry method.Results Vascular smooth muscle cells,which were obtained by this method,could grow fast and well,reaching 5×105 within five or six days,immunofluorescence staining showed that the purity was more than 85%,intracytoplasmic protein expression of α-SMA-was positive,the cell morphology was long spindle,they grew radially and wispily,and showed the typical ＂peak and valley＂ morphological appearance.Conclusion This new method could obtain considerable number of high-purity aortic smooth muscle cells in vitro.