文章摘要
居小兵,王宁宁,杨俊伟,王笑云,孙 彬, 邢昌赢.成人主动脉血管平滑肌细胞的体外培养[J].南京医科大学学报,2008,28(11):1429~1432
成人主动脉血管平滑肌细胞的体外培养
Method of human vascular smooth muscle cells culture in vitro
投稿时间:2008-05-29  
DOI:10.7655
中文关键词: 人主动脉  血管平滑肌细胞  平滑肌肌动蛋白-α
英文关键词: human aorta  vascular smooth muscle cells(VSMCs)  smooth muscle actin-α(α-SMA)
基金项目:国家自然科学基金(30700368),江苏省自然科学基金青年科技创新启动基金(BK2007609),江苏省卫生厅"135"重点学科基金资助
作者单位
居小兵 南京医科大学第一附属医院江苏 南京 210029 
王宁宁 南京医科大学第一附属医院肾内科,江苏 南京 210029 
杨俊伟 南京医科大学第二附属医院肾内科,江苏 南京 210011 
王笑云 南京医科大学第一附属医院肾内科,江苏 南京 210029 
孙 彬 南京医科大学第一附属医院肾内科,江苏 南京 210029 
邢昌赢 南京医科大学第一附属医院肾内科,江苏 南京 210029 
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中文摘要:
      目的:建立成人主动脉血管平滑肌细胞的体外培养方法?方法:无菌条件下分离出成人胸主动脉的平滑肌层,剪成1 mm × 1 mm × 1 mm的碎片,用组织块贴壁法,以含胎牛血清的DMEM培养基进行培养,生长融合的细胞用0.25%的胰酶消化?传代?观察培养细胞的形态学特点,并用免疫荧光的方法检测血管平滑肌肌动蛋白α(α-SMA)的表达?结果:在贴壁培养10~12天后,组织块的边缘开始有少量细胞爬出,呈长梭形,胞浆透亮?丰富;继续培养1~2周后组织块边缘融合,呈典型的“峰谷”样表现?消化传至3~8代,细胞的生长特性未见明显改变,免疫荧光显示细胞α-SMA的表达丰富?结论:组织块贴壁法培养人主动脉血管平滑肌细胞是一种稳定?可行的实验方法?
英文摘要:
      Objective:To establish the method of human vascular smooth muscle cells(hVSMCs)culture. Methods:Tissues were separated from segments of human adult aorta obtained at chest surgery. Small pieces of medial layer(about 1 mm3) were placed in flask and cultured in DMEM containing 20% heat-inactivated FCS and supplemented with sodium pyruvate(1 mmol/L),L-glutamine(1 mmol/L), penicillin(100 U/mL) and streptomycin(100 mg/mL). The cells were subcultured with 0.25% trypsin. Purity of cultures was assessed by positive immunostaining for ?琢-smooth muscle actin(?琢-SMA). Results:Two weeks later,the colonies formed by migrated cells from the explants were collected at confluence and routinely subcultured. Cells used in the experiments were between passages 3 and 8. Immunofluorescence staining reavealed that VSMCs displayed elongated in shape with enormous a-SMA expression. Conclusion:The method of aorta explants culture can be used to get the VSMCs in vitro.
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