尿路平滑肌细胞的培养和鉴定  

The culture and identification of urinary tract smooth muscle cells

作　　者：周昕[1] 耿红全 吴湘如 谢华 陆建华 蔡冰 徐卯生 薛皓亮 陈方 

机构地区：[1]上海市儿科医学研究所组织工程学研究室,200092 [2]上海市新华医院上海儿童医学中心小儿外科 [3]上海市新华医院上海儿童医学中心病理科

出　　处：《中华泌尿外科杂志》2003年第5期348-350,共3页Chinese Journal of Urology

基　　金：国家自然科学基金资助项目 (3 0 170 93 61) ;上海市教委重点科研基金资助项目 (99ZD0 6)

摘　　要：目的　建立尿路 (膀胱、输尿管、肾盂 )平滑肌细胞分离、培养、扩增的常规方法。　方法　人肾盂及输尿管标本各 1例、人膀胱标本 3例 ,猪输尿管标本 1例、猪膀胱标本 5例 ,采用胶原酶消化法从中分离尿路平滑肌细胞 ,在含 10 %胎牛血清的DMEM中培养 ,观察细胞形态及扩增情况 ,用免疫组化方法鉴定细胞类型特异性蛋白质α 肌动蛋白 (α actin)。　结果　尿路平滑肌细胞生长良好 ,呈现典型的平滑肌细胞形态。细胞经 6次传代仍扩增迅速 ,免疫组化染色显示α actin阳性。　结论　此培养方法简单、可靠 ,短期内可获得大量纯净的尿路平滑肌细胞。Objective To establish a method of isolating,culturing,proliferating urinary tract(nephropelvics,ureter and bladder)smooth muscle cells (SMC). Methods Urinary tract SMC were isolated from sample tissues (1 from human nephropelvics,1 from human ureter,3 from human bladders,5 from porcine bladders and 1 from porcine ureter) by collagenase digestion and were cultured in DMEM supplemented with fetal bovine serum.Morphology and expansion of the cells were observed.SMC specific protein (α-actin) was identified by immunohistochemical methods. Results The cells grew well and presented a typical morphological feature of SMC.They could be passaged 6 times without a remarkable decrease in rate of cell proliferation.Immunohistochemical staining showed that α-actin was positive. Conclusions This urinary tract SMC cultural method was simple,reliable and fruitful.

关 键 词：尿路平滑肌细胞 培养方法 胶原酶消化法 DMEM 免疫组织化学 Α-肌动蛋白 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]