【摘要】 目的探讨大鼠肾小管上皮细胞的原代培养方法。方法在原有肾小管细胞培养方法的基础上进行改良,采用机械研磨过筛、Ⅰ型胶原酶消化法分离出肾小管节段,在DMEM/F12[含5%胎牛血清(FBS)和胰岛素-转铁蛋白-硒]培养基中培养,并进行ZO-1免疫荧光染色鉴定。结果细胞培养至第3天完全贴壁,为多边鹅卵石样;免疫荧光染色显示90%以上的细胞为ZO-1表达阳性。结论采用机械研磨过筛、Ⅰ型胶原酶消化、DMEM/F12(含5%FBS和胰岛素-转铁蛋白-硒)培养基中培养的方法可以获得纯度达90%以上的肾小管上皮细胞,可满足对肾小管上皮细胞的研究。更多还原

【Abstract】 Objective To study the primary culture method of rat renal tubular epithelial cells.Methods On the base of original renal tubular epithelial cell culture method,the improvement was conducted.The renal tubular segments were isolated through mechanical grinding,sieving and collagenase Ⅰdigestion,then cultured with DMEM/F12 medium including 5%FBS and insulin-transferrin-selenium,under 5% CO2 at 37 ℃.The cultured cells were identified by fluorescent staining with ZO-1.Results The cells were cultured for 3 d and completely adhered.The cells were large,displaying the typical cobblestone appearance.Fluorescent staining showed that more than 90% cells were ZO-1 positive.Conclusion Mechanical grinding,sieving,collagenase Ⅰdigestion and culturing with 5%FBS DMEM/F12 medium plus insulin-transferrin-selenium may obtain the renal tubular epithelial cells with the purity of over 90% and may meet the study on renal tubular epithelial cells.更多还原