人尿路变移上皮种子细胞的体外培养与鉴定  

Culture in vitro and identification of human urinary transitional epithelial cells

作　　者：谷超[1] 李静[2] 隋志芳[2] 畅继武[2] 

机构地区：[1]天津医科大学基础医学院生物学教研室,天津300070 [2]天津医科大学第二医院泌尿外科研究所

出　　处：《山西医科大学学报》2010年第1期27-29,96,共4页Journal of Shanxi Medical University

基　　金：天津医科大学科学基金资助项目(2008KY04)

摘　　要：目的对人尿路变移上皮细胞进行原代和传代培养,探讨并建立细胞的培养体系,用于泌尿系统组织工程种子细胞的相关研究。方法采集人肾脏手术标本的肾盂、输尿管正常上皮组织利用组织块法,使用添加附加表皮生长因子(EGF)及牛垂体提取物(BPE)成分的角朊细胞无血清培养液(KSFM)进行细胞原代培养,用含EGF及BPE的KSFM进行传代培养。观察培养细胞的形态和增殖情况,经HE染色和免疫细胞化学SP法染色进行鉴定。结果培养制备的人尿路上皮细胞呈现典型的变移上皮细胞形态特征,原代培养9-12d时细胞汇合可达80%。免疫细胞化学染色显示细胞角蛋白(CKAE1/AE3)阳性。分离培养的为单一的人尿路变移上皮细胞,无成纤维细胞混杂。结论采用组织块法分离培养的人尿路变移上皮细胞在添加EGF及BPE的KSFM培养液中可维持良好的生物学特性,并具有一定的增殖传代能力。该培养体系所获得的细胞可用于人泌尿系统的基础研究及组织工程的进一步研究。Objective To explore the primary and second culture conditions of human transitional epithelial cells for further research on tissue engineering. Methods Cells of urothelium from the renal pelvis and ureter of human kidney surgery specimen were cultured by tissue explant methods with keratinocyte serum free medium（ KSFM ）with additional component, bovine pituitary extract （BPE）and epidermal growth factor（EGF）. The primary, cells were identified by cell morphology, HE and immunocytochemical SP staining. Results The cultured human urothelium cells possessed a typical morphological feature of transitional epithelial cells and their conflu- ence attached to 80% at day 9 - 12 after primary, culture. By immunocytochemistry, cultured cells showed cytokeratin AE1/AE3 （ CKAE1/AE3 ） positive. Cultured cells were pure human urinary transitional epithelial cells without fibroblasts. Conclusion Pure human urinary transitional epithelial cells can be cultured successfully by tissue explant method using KSFM with EGF and BPE, and retain the biological properties and capacity of proliferation. This culture method can be applied to the basic and further research on tissue en- gineering of human urinary system.

关 键 词：人尿路上皮 变移上皮细胞 细胞培养 组织工程 

分 类 号：Q813.1[生物学—生物工程]