[Isolation, primary culture and characterization of mouse glomerular mesangial cells].

To establish a method for the isolation, primary culture and characterization of mouse mesangial cells (MCs).We used two-layer micropore filter device to isolate glomeruli, and the MCs were cultured with eugenic selection method. Cell structures of MCs were observed by inverted phase contrast microscope, HE staining, scanning electron microscope and transmission electron microscope. Expressions of α-smooth muscle actin (α-SMA and nephrin were investigated by immunofluorescent cytochemistry. We used angiotensin II to stimulate MCs and observed the biological characteristics of the cells. The growth curve of MCs was examined by CCK-8 assay.After 20-30 days, the primary cultured MCs gradually covered the bottom of the dish. The mouse MCs usually attached to the surface 6-10 hours after passage, followed by the exponential phase in 2-3 days and the platform period in 3-5 days. Immunofluorescent staining showed that the expression of α-SMA was positive and nephrin was negative. MCs contraction was observed after the cells were stimulated by angiotensin II for 15 minutes.A method for the isolation, culture and characterization of mouse mesangial cells has been successfully established.