Genome wide analysis of Visceral adipose tissue CD14+ cells from obese subjects

Identification of the inflammatory signature in visceral adipose tissue CD14+ cells (adipose tissue macrophage) Total RNA obtained from CD14+ cells (Immunoselcted cells from stromal adipose tissue cells) Adipocytes and cells of the stroma vascular fraction (SVF) were obtained by collagenase digestion of adipose tissue. SVF cells were resuspended in endotoxin-free PBS supplemented with 2% FCS and 1 mM EDTA. Isolation of CD14+ using positive selection magnetic beads (Stemcell technologies) and CD4+ cells was performed using positive selection magnetic beads (Stemcell Technologies, Vancouver, Canada). An Illumina (San Diego, CA) RNA amplification kit (NuGEN, BiotinIL Module) was used according to the manufacturer's instructions to obtain biotinlabeled cDNA from 50 ng of total RNA extracted from adipose tissue CD14+ cells.