Endothelial cell growth medium ecm

Normal human astrocytes (NHAs), as a reference against the GBM cell lines, were provided by ScienCell Research Laboratories, Inc. (San Diego, CA, USA) and cultured in the provided astrocyte growth media supplemented with recombinant human epidermal growth factor, insulin, ascorbic acid, GA-1000, L-glutamine and 5% fetal bovine serum (FBS; SciencCell Research Laboratories, Inc.). U87, U251, T98, LN229 and U118 cells purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and were cultivated in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% FBS. The U87 cell line is a glioblastoma of unknown origin and the catalogue number of the cell line is ATCC HTB-14. In addition, the short tandem repeat profiling method was used to authenticate the U87 cells. ScienCell Research Laboratories, Inc. provided the human umbilical vein endothelial cells (HUVECs) and they were raised in endothelial cell growth medium (ECM) (ScienCell Research Laboratories, Inc.) supplemented with 1X endothelial cell growth supplement (ScienCell Research Laboratories, Inc.), 100 U/ml penicillin, 100 µg streptomycin/ml and 5% FBS. All cells were cultured at 37°C in 5% CO₂. Prior to each experiment, the cells were incubated for 24 or 48 h at 37°C in 1% O₂, 5% CO₂ and 94% N₂.