Rat proximal tubular epithelial cells in primary culture and identification.docVIP

 PAGE \* MERGEFORMAT 9 Rat proximal tubular epithelial cells in primary culture and identification Study: Fan Ying Zhang Lihong Chen Shuang Wang Guanglan quartz will love Sun Huajun Liu Shan Wu Abstract Objective To establish a better rat proximal renal tubular epithelial cells in primary culture model. Methods Wistar rats were obtained sterile kidney cortex was cut taken by collagenase digestion and 45% Percoll discontinuous density gradient centrifugation were purified with 10% FBS DMEM/F12 medium for primary culture and passaged, Cell morphology and by immunohistochemical staining and enzyme chemical staining. The results of 4 ~ 5 d cultured cell fusion into a single layer, showing typical cobblestone-like, cytokeratin 18 and were positive for alkaline phosphatase staining, cells can be passed 2 or 3 generations. Conclusion This method can be obtained in larger quantities in a short, reproducible proximal tubular epithelial cells, renal tubular cells in vitro provides an experimental platform for disease. Keywords: proximal tubular epithelial cells; primary culture; rat Abstract Objective To establish the better model for primary culture of rat proximal tubule epithelial cells (PTCs). Methods Wistar rats kidney were sterile taken out, then the cortex was separated and shredded, purified by collagenase digestion and 45% percoll discontinuous density gradient centrifugation, cultured and passaged by DMEM/F12 medium supplemented with 10% fetal bovine serum, identified by morphology, immunocytochemistry staining and enzyme staining.Results After 4 to 5 days, the cells were inosculated into a single layer and showed the typical cobblestone like appearance.The cytokeratin18 and alkaline phosphatase staining were positive, the cells could be passed 2 ~ 3 generation.Conclusions This method can yield large quantity and good repeatability PTCs in short term, those cells can provide an experimental platform for the research of pathological changes and