Gene expression analysis of gamma-M.tuberculosis-stimulated CD14hiCD16- and CD14loCD16+ monocytes

Current study will help us to elucidate the differential gene expression profiles of two different subsets of monocytes stimulated with gamma-M. tuberculosis CD14hiCD16- and CD14loCD16+ monocytes were isolated from human peripheral blood using magnetic beads for CD14 and CD16. 1.5 million cells were cultured in 1 ml of complete RPMI medium and stimulated with 10 microgram/ml of gamma-M. tuberculosis for 48 hours. After 48 hours cells were harvested and RNA was isolated from control and stimulated samples from CD14hiCD16- and CD14loCD16+ monocytes. So total we had 4 samples: 1. Control CD14hiCD16- 2. Stimulated CD14hiCD16- 3. Control CD14loCD16+ monocytes and 4. CD14loCD16+ monocytes. RNA from all these above 4 samples were sent to Phalanx Biotech Group for whole genome microarray.