Objective To find the optimal method of culturing primary podocytes in mice by comparing different glomerular separation techniques.Methods The renal glomeruli of C57/BL6J mice were obtained by sieving and with Dynabeads,while Bowman’s capsules were removed by digesting the glomeruli in type IV collagenase. Furthermore,the inverted phase contrast microscope was used to observe the structure of isolated renal glomeruli,expressions of podocin,while DAPI was observed by double-labeling immunofluorencence. Finally,the levels of podocin mRNA were measured by PCR.Results By using the conventional sieving method, the number of glomeruli isolated from one C57/BL6J mouse was 10 421±2421 with a purity of 90.2%±1.6%.The glomerular structure was complete and renal tubule fragments could be seen. However, with the Dynabeads method,the number of glomeruli isolated from one C57/BL6J mouse was 16112±2651 with a purity of 96.7%±1.2%,the glomerular structure was complete and there were few renal tubules. The purity and number of glomeruli obtained with Dynabeads were higher or larger than those obtained with the differential screening method,and the difference was statistically significant.A larger number of cobblestone-like cells that outgrew around nearly each glomerulus from the 7th to 10th day were observed with immunofluorescent staining, which was the characteristic of podocytes. The PCR results showed that the podocytes expressed podocin.Conclusions The test result has offered more proof that Dynabeads method is superior to the differential sieving method in isolating glomeruli from C57/BL6J mice because it is simple and highly effective for culturing primary glomerular podocytes.