Abstract:

BACKGROUND: An excellent primary culture of renal proximal tubular epithelial cells provides basis for drugs screening and mechanism research.
OBJECTIVE: To establish a modified method for the primary culture of mouse renal proximal tubule epithelial cells, and to test it urate uptake function.
METHODS: Kidneys were harvested and the cortices were dissected, minced, and digested using collagenase type Ⅰ. Fragments of the tissues were filtered with 50 and 100 mesh filter. Then the deposition was dissociated by gradient centrifugation with percoll. The isolate was implanted in the cell culture flasks. The bred cells were sub-cultured and identified by SP immunocytochemistry. Uric acid uptake function was tested by a range of probenecid and benzbromarone concentrations in transport medium contained 1 500 μmol/L uric acid.
RESULTS AND CONCLUSION: The adherent rate of segments of renal proximal tubules reached 54.5% after digested using 400U collagenase type I for 20 minutes; the most suitable time for the first change of the culture medium was 72 hours; Log phase began from the fifth day and the cell’s growth conditions become bad in the tenth day. In the transport medium contained    1 500 μmol/L urate acid, the 30-min uptake of uric acid was the highest and was taken as a measure of an initial rate. Different urate uptake inhibitions were appeared from different concentrations of the indicate drugs probenecid and benzbromarone. The inhibitions of benzbromarone were higher than probenecid. The mouse renal proximal tubular epithelial cells cultured by the modified method will yield rich and homogeneous harvest, and present well function of urate acid uptake. The study offers a model in vivo for pharmacological studies.