Human Cell Culture and Receptor Inhibitor Analysis

Human dermal microvascular endothelial cells (HMEC-1) and human embryonic kidney cells 293 (HEK293) were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HMEC-1 were cultured in MCDB131 medium (Gibco, Thermo Fisher Scientific Inc., Waltham, MA) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM l-glutamine (Gibco), 10 ng/mL epidermal growth factor (EGF, Sigma-Aldrich, St. Louis, MO) and 1 μg/mL hydrocortisone (Sigma-Aldrich). HEK293 cells were cultured in Dulbecco’s modified Eagle medium (DMEM; Gibco) supplemented with 10% FBS (Gibco), 100 U/mL penicillin and 100 μg/mL streptomycin sulfate (Gibco). Both cell types were cultured in a humidified atmosphere (5% CO₂, 37 °C), with medium changes twice a week. BMS-754807 (referred to as RI in this article), a selective small molecule receptor inhibitor of IGF-1R and InsR (Carboni et al., 2009 (link); Wittman et al., 2009 (link); Kolb et al., 2011 (link)), was obtained from Selleck (Houston, TX) and used at a concentration of 1 μM for in vitro assays.