Microarray analysis of mRNA expression in human microvascular endothelial cells exposed to colorectal cancer cell derived stimuli

Microarray analysis of mRNA expression in human microvascular endothelial cells exposed to colorectal cancer cell derived stimuli Human microvascular endothelial cells (HMECs) were seeded at 3x10˄5 cells in complete EGM2-MV growth medium on PET transwells of 1 µm pore size to form a confluent layer within 24 h and were then cultured for another 24 h in EGM2-MV medium including 5% FCS but no additional growth factor supplements. HT-29 colon carcinoma cells were seeded in the same growth medium at 4x10˄6 per 30 mm well and conditioned supernatant was collected from 24 to 48 h after cell seeding. HMEC cultures in transwells were left untreated or were stimulated by tumor-conditioned medium from the basal (experiment 1) or apical (experiment 2) side for 4 hours prior to isolation of total RNA.