Percoll法分离培养肾小管上皮细胞

目的建立良好的肾小管上皮细胞培养方法。方法选用SD幼鼠的肾组织进行细胞培养，采用Percoil密度梯度离心法分离纯化肾小管后，选择含10％新生牛血清的DMEM／F12培养基、5％CO2、37℃孵箱进行细胞培养。利用传1代的细胞，制作细胞爬片，用免疫组化、透射电镜进行鉴定。结果细胞生长4—5天后基本达到融合，细胞呈多边鹅卵石铺路样。结合形态学及免疫组化鉴定，细胞培养传1代后98％的细胞为肾小管上皮细胞。结论用Pereoll法分离培养的细胞数量多，均一性生长较好，可重复性操作较好。为体外研究接近组织原位RTECs细胞功能和特征及药物的筛选提供了可行性和可能性。

Dissociation and culture renal tubular epithelial cells by gradient centrifugation

Objective To establish a desirable culture method for rat renal proximal tubule cells(RTECs).Methods The kidneys of young SD rat were processed to separate and purify the renal tubule cells by Percoll gradient centrifugation.The cells were cultured in DMEM/F12 supplemented with 10% calf serum.The bred cells were subcultured and identitied by immunocytochemistry and transmission electron microscopy.Results After 4-5 days,the cultured cells reached almost complete confluence,displaying the typical cobblestone layout.According to the results of cell morphology and immunocytochemistry,the purity of secondary cultured renal tubular epithelial cells reached 98%.Conclusion The RTECs dissociated by Percoll gradient centrifugation,when cultured,will yield rich and homogeneous harvest,offering a model,simulating the RTECs in vivo,for physiological and pharmacological studies.