肾小管上皮细胞原代培养及鉴定

Primary culture and identify of renal tubular epithelial cells

目的探讨肾小管上皮细胞体外培养及鉴定方法。方法①采用机械分离结合酶消化法获取肾小管,原代培养肾小管上皮细胞。②利用动态观察、细胞化学染色法及细胞的透射电镜扫描进行鉴定。结果①肾小管段不同换液时间24h、48h、72h、96h、120h,贴壁率分别为:0.1378±2.048E-02、0.3300±7.969E-02、0.5411±3.919E-02、0.5344±2.506E-02、0.4389±3.018E-02,以72小时首次换液最佳。②肾小管上皮细胞成功原代培养并于第4～7天处于对数生长期。细胞碱性磷酸酶染色均呈阳性,透射电镜扫描见细胞面向液层的刷状缘有大量微绒毛形成。结论采用机械结合酶消化法可分离到较纯的肾小管,肾小管段培养72h首次换液细胞贴壁最佳,肾小管上皮细胞用化学结合形态学及鉴定。

Objective To investigate the method of primary culture and identify of renal tubular epithelial cells. Methods ①The renal tubular segments were collected by mechanical and chemical digestive method and the renal tubular epithelial cells were primarily cultured. ②The renal tubular epithelial cells were identified by dynamic observation, cytochemistry staining, and transmission electronic microscope. Results ①The anchor rates of renal tubular at 24,48,72,96 and 120 hours were 0.1378±2.048E-02,0.3300±7.969E-02,0.5411±3.919E-02,0.5344±2.506E-02,0.4389±3.018E-02 respectively and the optimum time of the first replacing medium was 72 hours. The primary culture of the renal tubular epithelial cells succeeded and cells grew in logarithm growing phase from 4 to 7 days. ②The primary cultured cells were identified successfully. The cultured cells were all positive by alkali-phosphatase staining and under transmission electron microscopy the brush border of the renal tubular epithelial cells had a great many of microvilli. Conclusion The machanical and chemical digest method can obtain fairy pure renal tubular segments. The growth time is the third day after the first replacing medium. The renal tubular epithelial cells can be cultured successfully with our culture method.