家兔主动脉平滑肌细胞原代培养的研究  

A new method for culturing vascular smooth muscle cells from the rabbit aorta

作　　者：王伟[1,2] 戴敏[1] 潘礼龙[1] 

机构地区：[1]安徽中医学院药学院中药药理学教研室·现代中药安徽省重点实验室,安徽合肥230038 [2]合肥师范学院生物系,安徽合肥230061

出　　处：《中国药理学通报》2008年第11期1534-1537,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30772748);安徽省优秀青年科技基金资助项目(No0804016905)

摘　　要：目的探索高效快速的家兔血管平滑肌细胞体外原代培养技术,为分子生物学实验提供足量可靠的细胞来源。方法取家兔主动脉中层,用刀片切成约1mm3,采用不同浓度的Ⅰ型胶原酶以及不同消化时间对组织块进行预消化,然后将组织块贴在培养瓶上进行培养。结果1.5g.L-1胶原酶消化6h后组织块细胞迁出速度及组织迁出率明显提高(P<0.05),组织块贴壁后24h即可见血管平滑肌细胞从组织块周围游离出来,呈梭形或星形,d4～5即可进行传代培养,传代后d3～4即可融合成致密单层细胞,呈典型的峰谷交错状,经免疫组化SP法鉴定有α-actin的表达,进一步确认其为血管平滑肌细胞。结论经1.5g.L-1胶原酶消化6h后组织块细胞迁出速度且迁出率明显提高,从而大大缩短原代培养的时间。Aim To develop a convenient and effective method to isolate and culture primary rabbit aortic vascular smooth muscle cells （VSMCs）. Methods The thoracic aortas were removed by dissection under sterile conditions. Aortic smooth muscle cells were excised and cleaned of fat and connective tissue, and the isolated vessel media was cut into 1 mm3 pieces. The explants were digested with different concentrations of collagenase type 1 ,and incubated at 37℃ for different time,then undispersed explants were placed onto a sterile 100-mm plastic tissue culture dish with growth medium. Results VSMCs could emi- grate from the explants digested 6 h by collagenase type [ （ 1.5 g · L^-1 ） for 24 hours, the cells would passage for another 4 - 5 days. Confluency could be reached within 3 N 4 days after subculturing. VSMCs were identified by immunoreactivity with α-actin and by the smooth muscle cell-specific, hills and valley-like morphology. Conclusion It was an effective method to culture primary VSMCs from the explants digested for 6h by collagenase type I （1.5 g · L^-1） , which could shorten primary culture time.

关 键 词：兔 主动脉 血管平滑肌细胞 原代培养 

分 类 号：R-332[医药卫生] R322.121