维生素C对大鼠肾小管上皮细胞增殖活力的影响  

Impact analysis of Vitemin C on the proliferating ability of rat renal tubular epithelial cell

作　　者：陈昕[1,2] 赵雷[1,3] 周余来[4] 

机构地区：[1]暨南大学第二临床医学院 [2]深圳市人民医院检验科,广东深圳518020 [3]深圳市人民医院麻醉科,广东深圳518020 [4]吉林大学药学院生物工程教研室,吉林长春130021

出　　处：《中国实验诊断学》2013年第3期452-454,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的优化大鼠肾小管上皮细胞的培养方法,为肾脏疾病的体外研究提供一定的技术支持。方法采用Ⅱ型胶原酶消化法结合Percoll密度梯度离心法,分离纯化肾小管节段,对肾小管节段进行贴壁培养,待上皮细胞爬出后,通过流式细胞仪分析细胞角质素-18(CK-18)的表达情况,并用免疫组化方法对CK-18在肾小管上皮细胞中的表达进行定位;通过不同剂量的维生素C作用,CCK-8方法分析大鼠肾小管上皮细胞的增殖活力改变。结果肾小管节段贴壁培养1-2天后,肾小管上皮细胞从节段内爬出,培养3-4天,细胞密集生长;随着传代进行,肾小管上皮细胞凋亡比例增加,增殖活力减弱,在维生素C的作用下,可在一定程度上减缓细胞的老化速度。结论维生素C可适度延长肾小管上皮细胞的体外培养时间,增强细胞活力,方法经济简便。Objective Refine the culture method of rat renal tubular epithelial cells （RTECs） ,so as to provide tech- nique support for the research in vitro of kidney disease. Methods The renal tubules were separated with the digestion of II type collagenase and purified with Percoll density gradient centrifugation. After the RTECs grown out from the tu- bule,cytokeratin-18 （CK-18） was detected with flow cytometry and located with immunohistochemistry. The ability to proliferate of RTECs, under effect of Vitemin E in each dose, was evaluated with CCK-8. Results Cultured for 1-2 days,RTECs began to grow outside from the tubule and growed imtensively after cultured for 3-4days. The RTECs were dead gradually,with more weaken ability to proliferate while passage. Under the effect of Vitmin C, slower speed to apolexis was achieved in a degree. Conclusion The culture ＇time could be extended by Vitemin C,with better ability to proliferate, and the method is cheap as well as convenient.

关 键 词：肾小管上皮细胞 维生素C 增殖活力 

分 类 号：R692[医药卫生—泌尿科学]