改良成人大隐静脉平滑肌细胞培养方法  

Research of Human Saphenous Vascular Smooth Muscle Cells Culture in Vitro

作　　者：刘朝兵[1] 王志维[1] 吴红兵[1] 夏军[1] 

机构地区：[1]武汉大学人民医院心血管外科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2009年第4期496-497,521,I0003,共4页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:30872537)

摘　　要：目的:研究提高成人大隐静脉平滑肌细胞(hVSMC)体外培养效率的方法。方法:无菌条件下取3 cm左右大隐静脉,去除内外膜后,切成1 mm×1 mm大小组织块,贴块法培养,高糖DMEM中加入血小板衍生生长因子(PDGF)10μg/L,免疫组织化学法鉴定平滑肌细胞。结果:大隐静脉平滑肌细胞沿组织块长出时间为8-12 d,原代培养(22±2)d传第1代,免疫组织化学染色细胞平均阳性率≥98%。结论:PDGF和高糖克服了hsVSMC体外培养休眠期过长的缺点。运用该方法,稳定获得hsVSMC原代细胞,且重复性好,细胞产量较大,简便而有效。Objective： To study on increasing the successful rate of in vitro culturing of the human saphenous vascular smooth muscle cells （hVSMCs）. Methods： A group of 3-cm-long saphenous veins were harvested under sterile conditions. After the internal and external membrane were removed, the middle membranes were cut into 1 mm× 1 mm size pieces, and were cultured by tissue-piece inoculation method, added to DMEM medium with PDGF （10 μg/L） and high concentration of glucose. The smooth muscle cells were identified by immunohistochemistry. Results： The hVSMCs grew along the tissue pieces for 8-12 days, the original generation was cultured for （22±2）days, and the purity of the third passage SMCs was more than 98%. The cultured cells showed the typical peak and valley appearance under microscope. Immunohistochemistry staining with m-Ab against rabbit SM-α-actin identified the cells. Conclusion： Culturing with PDGF and high glucose in vitro overcomes the shortcoming of the long period of hsVSMC dormancy. And the method was reproducible, simple, stable, and effective.

关 键 词：大隐静脉 平滑肌细胞 细胞培养 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学]