Mesangial Cell Culture Conditions

Human renal mesangial cells (HRMCs) were purchased from ScienCell Research (San Diego, United States), and the mouse mesangial cell (MMC) line SV40 MES-13 was purchased from the American Type Culture Collection (ATCC). All cells were cultured in DMEM (Gibco, United States) containing 10% fetal bovine serum (Gibco, Australia), streptomycin (100 μg/mL) and penicillin (100 U/mL) at 37°C and 5% CO₂. Standard medium containing 5 mM D-glucose was used for the normal glucose (NG) condition. We added 20 mM glucose (yielding a final concentration of 25 mM) or mannitol for the high glucose (HG) condition or osmotic control (MA) and stimulated the cells for 48 h. The concentration was chosen based on our previous studies (Lu et al., 2015 (link); Wang et al., 2018a (link); Ma et al., 2019 (link)).