〔Abstract〕 Objective To explore a simple, convenient and reproducible method for thein vitroextraction, culture and identification of primary glomerular mesangial cells(GMCs) from non-human primate. Methods The kidneys of macaca mulattas were removed under aseptic conditions, and the glomeruli were separated using two different pore sizes(between 100 and 200 mesh) and digested with type VI collagenase at different concentrations(0.1 mg/ml and 0.2 mg/ml), and different digestion times(10 min and 15 min) were set for the two concentrations. The growth of GMCs under different digestion conditions was observed in the bottles. The structure of GMCs was observed under microscope and the expression of α-smooth muscle actin(α-SMA) and Nephrin in GMCs was detected by immunofluorescence and Western blot. The biological properties of GMCs were observed after stimulation with tumor necrosis factor-α(TNF-α), and the viability and proliferation of GMCs were detected by CCK-8 and High Connotation Cell Imaging System, respectively andthe migration ability of GMCs was detected by Transwell and cell scratch assay. Results The glomeruli collected using 0.1 mg/ml type VI collagenase digestion for 10 min were more apposed and the glomeruli cells were in a better growth state. Glomeruli in primary culture were adherent at 3 d. Irregularly shaped or star-shaped cells began to migrate at 7 d. GMCs gradually spread to the bottom of the bottle after 21～35 d. GMCs were positive for α-SMA protein and negative for Nephrin protein. 10 ng/ml TNF-α in vitro stimulation significantly enhanced the proliferation and migration ability of GMCs. Conclusion The method of collagenase digestion(0.1 mg/ml type VI collagenase digestion for 10 min) was successfully established for the isolation and culture of GMCs, which can provide a suitable cell model to better simulate human kidney diseases in the future.