SD rat renal tubular epithelial cells in primary culture and identification.docVIP

 PAGE \* MERGEFORMAT 15 SD rat renal tubular epithelial cells in primary culture and identification OF: He Jin Yuan, Tsang Kwok Wa, Wu, the Zhong Wen, Gui Zhiming, like Michael Lin [Abstract] Objective: To investigate the renal tubular epithelial cells in vitro and identification method for experimental kidney disease research platform. Methods: mechanical grinding, isolated by collagenase digestion and renal tubular segments containing 10% fetal bovine serum and 1% of epithelial cell growth factor in epithelial cells grown in culture, 0.25% trypsin digestion subculture, and use primary and passage 1 cells for immunocytochemical staining. Results: The cell culture to 3 days completely adherent, 4 ~ 7 d in the logarithmic phase for multilateral cobblestone; immunocytochemistry showed positive expression of cytokeratin 18. Conclusion: mechanical grinding, collagenase digestion method combined with cell culture medium can have a more pure tubular, renal tubular epithelial cells are cultured ideal method for further study of the causes and mechanisms of urinary stones provide an experimental foundation. [Keywords:] renal tubular epithelial cells; primary culture; CK18; rat [Abstract] Objective: To discuss the method of the primary culture and identification of SD rat s renal tubular epithelial cells, thus to supply an experimental platform for the study of renal calculi.Methods: The renal tubular segments were isolated by mechanical grinding and collagenase digestion. The collected cells were cultured with EpiCM, which contained 10% FBS and 1% EpicGs, in incubation with 5% CO2 at 37 , and subcultured with 0.25% trypsin.The cultured cells were identified by immunocytochemistry. Results: The cultured cells were completely adhered after 3 days, the logarithmic growth phase was beween 4 and 7 days. The cells were large, displaying the typical cobblestone appearance.Immunocytochemistry suggested that the expressions of cytokeratin 18 in renal tubular epi