输尿管上皮细胞及平滑肌细胞原代培养方法的比较  

Primary culture of ureteral epithelial and smooth muscle cells culture in vitro

作　　者：宋南[1] 符伟军[1] 王忠新[1] 李钢[2] 王晓雄[3] 张旭[3] 刘振湘[4] 白志明[4] 

机构地区：[1] 解放军总医院海南分院泌尿外科,三亚572014 [2] 解放军第三○九医院全军器官移植研究所 [3] 解放军总医院泌尿外科 [4] 海口市人民医院中南大学湘雅医学院附属海口医院

出　　处：《中华实验外科杂志》2014年第9期1974-1976,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81070555）;北京市自然科学基金资助项目（7142145）;军队临床高新技术重点资助项目（413DG63J）

摘　　要：目的 比较组织块法与酶消化法两种不同细胞原代培养方法,探讨更有效的输尿管上皮细胞及平滑肌细胞的体外培养方法.方法 取临床肾癌切除术后患者的1段输尿管,分成2组,分别利用组织块法及酶消化法进行原代培养,比较两种方法培养的尿路上皮细胞及平滑肌细胞的成功率以及细胞长出所需平均时间.通过倒置显微镜、免疫组织化学染色法进行原代培养细胞的鉴定.结果 应用组织块法细胞原代培养成功率明显高于酶消化法.输尿管上皮细胞组织块法成功率为86.7％,细胞长出所需时间为（6.25±1.50）d;酶消化法成功率为20.0％,细胞长出所需时间为（3.15 ±0.80）d,两种方法获得的细胞均符合正常输尿管上皮的细胞形态及生物学特征,生长良好.输尿管平滑肌细胞组织块法成功率为66.7％,细胞长出所需时间为（5.05±1.50）d;酶消化法成功率为13.3％,细胞长出所需时间为（2.00±1.20）d,两种方法获得的细胞均符合正常输尿管平滑肌细胞形态及生物学特征,生长良好.结论 组织块法原代培养输尿管上皮细胞及平滑肌细胞培养成功率明显高于酶消化法,酶消化法细胞长出时间明显短于组织块法.Objective To compare the tissue piece method and tissue enzyme digestion method for primary cell culture of human epithelial and smooth muscle cells from the ureter in vitro.Methods The ureter specimens were taken from patients subject to clinical resection of renal carcinoma of kidney,and divided into two groups：tissue piece method group and tissue enzyme digestion method group.Indexes （success rate and average days for cells growth） were analyzed to evaluate efficiency of the two methods.Primary cultured cells were identified by inverted microscope and immunohistochemical staining.Results Success rate of tissue piece method was significantly higher than that of enzyme digestion method.For tissue piece method,success rate of tissue piece method was 86.7% （ureteral epithelial cells） and 66.7% （ureteral smooth muscle cells）,and the average days for cells growth was （6.25 ± 1.50） days （ureteral epithelial cells） and （5.05 ± 1.50） days （ureteral smooth muscle cells）.For enzymatic digestion method,success rate was 20.0% （ureteral epithelial cells） and 13.3% （ureteral smooth muscle cells）,and the average days for cells growth was （3.15 ±0.80） day （ureteral epithelial cells） and （2.00 ± 1.20） days （ureteral smooth muscle cells）.Morphology and biological characteristics of the cells were in line with the normal ureter epithelial and smooth muscle cells.Conclusion For the primary culture of ureteral epithelial cells and smooth muscle cells,success rate of tissue piece method is significantly higher than that of enzymatic digestion method,and the time for cells growth is shorter in enzyme digestion method than in tissue piece method.

关 键 词：原代培养 酶消化法 输尿管上皮细胞 输尿管平滑肌细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]