Gene expression of Myeloid cells stimulated with cytokines.

THP-1 is a human monocytic cell line derived from an acute monocytic leukemia patient. It is efficiently converted into macrophages using cytokine combinations and or by using chemical compound Phorbol 12-myristate 13-acetate (PMA). Together with performing gene expression analysis on HSC’s we also performed gene expression analysis on THP-1 cells in order to check which genes have been expressed/overexpressed during differentiation into macrophages. Cells were given treatment with cytokines combination (GMCSF, MCSF). After 24 hours of incubation, cells were washed and pellet was preserved in RNAlater (Ambion) to be shipped to Genotypic Technologies Pvt. Ltd. (Bangalore) for microarray analysis. RNA was isolated by Qiagen’s RNeasy mini kit and was reverse transcribed into cDNA. Cy3 labeled cRNA was formed by cDNA by T7 promoter based linear amplification using Agilent’s Quick amp labeling kit and quantified in Nanodrop Spectrophotometer and their integrity was checked by Bioanalyser. . THP-1 cells were treated with cytokines(GMCSF, MCSF) for 24hrs. After 24 hours of incubation, cells were washed and pellet was preserved in RNAlater (Ambion) to be shipped to Genotypic Technologies Pvt. Ltd. (Bangalore) for microarray analysis.