Gene expression profiling of CD34+ cells from MDS patients and normal controls

In order to gain insight into the poorly understood pathophysiology of the myelodysplastic syndromes (MDS), we have determined the gene expression profiles of the CD34+ cells of 55 MDS patients using the Affymetrix GeneChip U133 Plus2.0 platform Keywords: Disease v normal 55 patients with MDS and 11 healthy controls were included in the study. At the time of investigation, 18 patients had RA, 19 RARS and 18 RAEB. The RAEB group was further subdivided into RAEB1 and RAEB2 for a selected part of the analysis. Bone marrow samples were obtained and CD34+ cells isolated from MDS patients and healthy controls using MACS magnetic cell separation columns. Total RNA was extracted using TRIZOL following the protocol supplied by the manufacturer. RNA was amplified using the Two-Cycle cDNA Synthesis and the Two-Cycle Target Labelling and Control Reagent packages (Affymetrix). Samples were hybridized to GeneChip Human Genome U133 Plus 2.0 arrays. Affymetrix CEL files were pre-processed using Robust MultiChip Analysis (RMA).