文档介绍：Percoll法分离培养肾小管上皮细胞
【关键词】 Percoll
摘要: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Per-coll密度梯度离心法分离纯化肾小管后,选择含10%、5%CO 2 、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Percoll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。
关键词:近端肾小管上皮细胞;细胞培养;离心法;密度梯度
Abstract: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 cells were cultured in with10%calf bred cells were subcultured and identitied by immu-nocytochemistry and transmission electron After4-5days
,the cultured cells reached plete confluence,displaying the typical cobblestone to the results of cell morphology and immunocytochemistry,the purity of secondary cultured renal
tubular epithelial cells reached98%.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.
Key words:proximal renal tubular epithelial cell;cell culture;centrifugation;density gradient
近年来,在对各种肾缺血性损伤导致肾衰的研究中发现,尽管缺血性肾衰的发展有多种因素并可分为数个进展阶段,但