兔原代胸主动脉平滑肌细胞培养方法及不同浓度血清培养基对其增殖的研究  

Culture in different serum of vascular smooth muscle cells from rabbit thoracic artery

作　　者：刘苏健[1] 申庆民 吴继东[1] 刘宏[1] 赵京[1] 

机构地区：[1]北京航天总医院血管外科,100076

出　　处：《中国现代医药杂志》2013年第9期21-23,共3页Modern Medicine Journal of China

摘　　要：目的探讨体外分离培养兔原代胸主动脉平滑肌细胞（vascular smooth muscle cells, VSMCs）技术及在不同浓度血清培养基下生长情况。方法采用组织贴块法进行原代培养，分别用不同浓度血清培养基（10％、15％、20％）对原代VSMCs进行培养．胰酶消化法传代，用倒置显微镜对原代培养兔平滑肌细胞进行形态学观察并鉴定，MTT法测定增殖能力。结果体积分数20％血清的DMEM培养液培养的原代细胞生长出现明显对数期。培养5代的兔平滑肌细胞纯度达97％以上：镜下可见平滑肌细胞生长，免疫组化提示平滑肌细胞肌动蛋白阳性表达，细胞生长第4～5d内光密度值变化较明显。结论本法能有效提高兔动脉平滑肌细胞原代培养的成功率，为研究血管平滑肌细胞生物学行为提供了有效模型。Objective To observe the method that culture and identify vascular smooth muscle cells（VSMCs） from rab- bit thoracic artery. Methods The rabbit thoracic artery cell was cultured by substrate-attached method and the effects of dif- ferent serum content on growth were observed. The cultured cells were observed and identified by immunohistochemical methods through smooth muscle a-actin （a-SM-actin）, the proliferation ability were studied by MTT assay. Results Containing 20% serum DMEM high glucose culture medium of cells had an effect on apparent logarithmic phase.The purity of VSMCs were more than 97%. The passaged VSMC was expressed by the smooth muscle specific differentiation marker ot-SM-actin, the significant change of optical density was detected after culture for 4 to 5d. Conclusion The method is an effective model to culture smooth muscle cells for vascular surgery researches.

关 键 词：兔 胸主动脉 平滑肌细胞 原代培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]