人近端肾小管上皮细胞的原代培养  

Primary Tissue Culture of Proximal Tubular Epithelial Cells in Human

作　　者：严玉澄[1] 钱家麒[1] 戴慧莉[1] 许雁萍[1] 

机构地区：[1]上海第二医科大学仁济医院肾脏科,上海200001

出　　处：《上海第二医科大学学报》2005年第4期388-391,共4页Acta Universitatis Medicinalis Secondae Shanghai

摘　　要：目的应用组织块培养法建立人近端肾小管上皮细胞的原代培养方法。方法取肾皮质组织块,应用无血清培养液进行原代培养,融合后用胰蛋白酶-EDTA消化传代。取第三代细胞检测细胞角蛋白、波形蛋白、碱性磷酸酶、结蛋白、Ⅷ因子、α-肌动蛋白的表达,应用扫描电镜和透射电镜观察细胞的超微结构。结果融合的上皮单层细胞呈铺路石样表现,伴圆顶(dome)形成。细胞角蛋白、波形蛋白和碱性磷酸酶染色阳性,α-肌动蛋白、Ⅷ因子相关抗原、纤维连接蛋白和结蛋白染色阴性。细胞存在极性,细胞尖端存在短的微绒毛,细胞之间连接紧密。结论应用组织块培养法结合无血清培养液可以成功建立原代人近端肾小管上皮细胞系。Objective To establish primary tissue culture method of proximal tubular epithelial cells in human. Methods Kidney cortex explanted into the serum-free culture medium gave rise to a primary culture of kidney epithelial cells. After confluence, cells were subcultured using Trypsin-EDTA digestion. Cytokeratin, vimentin, alkaline phosphatase, α-actin, Ⅷ Ag, fibronectin and desmine were determined using the third generation cells. The ultrastructure were examinated by the transmitted and scanning electronic microscope. Results The confluent epithelial cell monolayer showed cobblestone appearance and demonstrated dome formations. The histochemistry staining of cytokeratin, vimentin, alkaline phosphatase were positive and that of α-actin, Ⅷ Ag, fibronectin and desmine were negative. There are microvilli on the cell surface and tight connections between the adjacent cells. Conclusion Primary tissue culture of proximal tubular epithelial cells combined with serum-free culture medium can be established in human.

关 键 词：细胞培养 近端肾小管上皮细胞 无血清培养液 细胞角蛋白 波形蛋白 碱性磷酸酶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]