鼠结肠平滑肌细胞的分离、培养与鉴定  

Isolation,culture and identification of smooth muscle cells from rat colon

作　　者：梁宁霞[1] 衣兰娟[1] 田琳[1] 刘翠萍[1] 魏睦新[1] 朱兰兰[1] 

机构地区：[1]南京医科大学第一附属医院中医科,210029

出　　处：《江苏医药》2005年第6期433-435,i002,共4页Jiangsu Medical Journal

基　　金：省科技厅国际合作项目(BZ2002066);江苏省135工程重点医学人才基金资助(135038)

摘　　要：目的建立体外培养鼠结肠平滑肌细胞的方法。方法应用酶解法分离大鼠的结肠平滑肌细胞,用含10%胎牛血清的DMEM液进行鼠结肠平滑肌细胞的原代培养及传代,并以免疫细胞化学对其进行鉴定。结果新鲜分离的结肠平滑肌细胞呈梭形,核居中。培养24h后细胞开始贴壁,3～5d后开始增殖,14d后细胞密集,呈峰谷样生长。细胞经平滑肌特异性肌动蛋白αactin鉴定,确定为平滑肌细胞。结论应用酶解法可得到急性分离的鼠结肠平滑肌细胞,用含10%胎牛血清的DMEM液重悬分离得到的结肠平滑肌细胞,经过10d左右的培养可以获得结肠平滑肌细胞。该方法重复性好,效果稳定。Objective To establish an efficient culture method of smooth muscle cells (SMCs) from rat colon in vitro.Methods Freshly isolated SMCs from the muscle layer of the distal colon were prepared by collagenase digestion,and cultured in DMEM supplemented with 10% fetal bovine serum. The cells were subcultured and identified by immunocytochemical staining.Results Freshly isolated SMCs were spindle-shaped with centrally located nuclei. In primary culture, SMCs attached to the culture vessels by 24h, proliferated by 48h, and reached confluency after 14 days with a 'hill-and-valley' pattern. Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Conclusion The procedure mentioned above is efficient and reproducible for isolation,culture and identification of rat colon smooth muscle cells.

关 键 词：结肠平滑肌细胞 培养与鉴定 免疫细胞化学 DMEM 胎牛血清 方法应用 原代培养 肌动蛋白 急性分离 酶解法 培养鼠 24h 特异性 重复性 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R595.7[医药卫生—基础医学]