Transcriptomic Alterations of Human Microvascular Lung Endothelial Cells in Response to TNF-α and Pathological Cyclic Stretch
Human microvascular lung endothelial cells (HMVLECs) purchased from Lonza Ltd. (Walkersville, MD) were cultured in the manufacturer’s recommended endothelial cell basal medium (EBM-2) media containing 10% fetal bovine serum (Clonetics, Walkersville, MD). Cells were grown at 37°C in a 5% CO2 incubator. Cells were initially grown in complete culture medium containing 10% fetal bovine serum, and media was changed every other day until cells reached confluence. All cyclic stretch (CS) experiments were performed with the FX-5000 Flexcell Tension Plus system (Flexcell International Co., Hillsborough, NC). On the day before the CS challenge, the cells were incubated in EBM-2 supplemented with 2% FBS, vascular endothelial growth factor, fibroblast growth factor, epidermal growth factor, hydrocortisone, ascorbic acid, insulin like growth factor-1, GA1000 (gentamycin/amphotericin-B).