动脉粥样硬化模型小鼠血管平滑肌细胞原代培养及鉴定

摘要

Objective : To set up a method for isolation and characterization of smooth muscle cell from aorta of atherosclerotic mouse model. Methods : The medium layer of aorta from ApoE-/- mouse was isolated, and primary vascular smooth muscle cells( VSMCs ) were obtained using tissue explant method characterized by immunohistochemical and immunofluorescent staining. Results : VSMCs migrated from explants of mouse aorta tissue after 3 days of culture. Two weeks later,VSMCs grew to form a fusing monolayer in a pattern of spiking and fading. Immunohistochemical and immunofluorescent staining demonstrated that the purity of VSMCs was more than 98％ . Conclusion:The tissue explant method is a simple, stable, and reliable method for the primary culture of VSMCs, which could be widely used to isolate VSMCs with high purity for in vitro functional tests.%目的:介绍动脉粥样硬化模型小鼠血管平滑肌细胞(VSMC)原代培养及鉴定的方法.方法:分离载脂蛋白E基因敲除小鼠主动脉,组织贴块法获得原代平滑肌细胞,免疫荧光及免疫组化方法检测细胞的纯度和分化状态.结果:培养第3天时,可见自组织块周围长出少量梭形或长梭形细胞,至培养2周细胞融合成片,呈"峰、谷"状生长.肌动蛋白免疫组化及荧光染色显示,细胞传至第6代后纯度在98%以上,证实应用此方法分离原代VSMC可以满足平滑肌体外功能实验的需求.结论:应用组织贴块法培养小鼠VSMC,操作简单,结果稳定,纯度较高,具有推广应用价值.