Hrmcs

Primary human renal mesangial cells (HRMCs) were purchased from ScienCell Research Laboratories (cat. 4200) and cultured in mesangial cell medium (MCM, cat. 4201) supplemented with 10% FBS (cat. 0010), 5 mL of mesangial cell growth factor (MsCGS, cat. 4252) and 5 mL of penicillin/streptomycin (P/S) solution (cat. 0503). Primary human glomerular endothelial cells (HRGECs) were purchased from ScienCell Research Laboratories (cat. 4000) and cultured in endothelial cell medium (ECM, cat. 1001) supplemented with 10% FBS, 5 mL of endothelial cell growth factor (ECGS, cat. 1052) and 5 mL of P/S solution. Cells were all maintained in 5% CO₂ at 37°C. PDGF‐BB (Peprotech, cat. 100‐14b, 20 ng/mL) and VEGFA (Peprotech, cat. 100‐20, 50 ng/mL) were used as irritants of MCs and ECs, respectively.

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)).

HRMCs were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA), and maintained in the Mesangial cell medium (ScienCell Research Laboratories) in a humidified atmosphere containing 5 % CO₂ at 37 °C. HRMCs were used between passages 3 and 6.