Gelatin coated cell culture plastic

Mouse embryonic fibroblasts were purchased from ATCC and cultured in DMEM, (Gibco) supplemented with 10% FBS and 1% Pen/Strep, (Sigma) until passage 4 – 6 on gelatin coated cell-culture plastic (BD Biosciences)

Primary human umbilical vein endothelial cells (HUVECs) were purchased from Lonza and expanded in EBM2 endothelial cell media (Lonza) supplemented with EGM-2 bulletkit until passage 6 – 10 on gelatin coated cell-culture plastic (BD Biosciences).

Mouse skeletal myoblasts (C2C12) were purchased from ATCC and expanded in DMEM (Gibco) supplemented with 10% FBS, and 1% HyClone Antibiotic Antimycotic solution (Thermo Scientific) on gelatin coated cell-culture plastic (BD Biosciences). Cells were cultured under standard culture conditions (37°C in 5% CO2). Medium changed every o ther day. To avoid myotube formation, cells were passaged with 0.05% trypsin/EDTA (cellgro-25052CI) at 70 – 80% confluency. For C2C12 differentiation, we supplemented DMEM (Gibco) with 2% horse serum (Gibco) and 1% antibiotic antimycotic solution (10,000 units/ml penicillin, 10,000μg/ml streptomycin, and 25μg/ml Amphotericin B; HyClone). C2C12 were expanded until passage 6 – 10.