Project description:Human PBMC, there are major T, B, Dc, Basophil, Eosinophil and Neutrophil subsets with typical biological functions. The characteristic behavior of each subset was determined by its unique gene expression and regulation. Difference between each subset or different treatment to the same subset will induce huge amount different gene expression. This will provide the basic evidence of understanding why and how each subset cells functions properly. We used microarrays to detail the global programme of gene expression in each subsets of human PBMC and identified distinctly up-regulated genes during this process.
Project description:This SuperSeries is composed of the following subset Series: GSE10512: Gene Expression Profiles of PBMC Subsets in Acute GVHD Following Cord Blood Transplantation (IMS v.1) GSE10513: Gene Expression Profiles of PBMC Subsets in Acute GVHD Following Cord Blood Transplantation (IMS v.2) Keywords: SuperSeries Refer to individual Series
Project description:High quality genetic material is an essential pre-requisite when analyzing gene expression using microarray technology. Peripheral blood mononuclear cells (PBMC) are frequently used for genomic analyses, but several factors can affect the integrity of nucleic acids prior to their extraction, including the methods of PBMC collection and isolation. In this study, we compared the Ficoll-Paque density gradient centrifugation and BD Vacutainer cell preparation tube (CPT) protocols to determine if either method offered a distinct advantage in preparation of PBMC-derived immune cell subsets for their use in gene expression analysis. We compared gene expression in PBMC and individual immune cell types from Ficoll and CPT isolation protocols using Affymetrix microarrays. PBMC from 3 healthy donors were isolated using Ficoll-Paque gradient or BD Vacutainer CPT methods. Subsequently, immune cell subsets were separated by positive selection from PBMC obtained by both methods. RNA was extracted from total PBMC isolated by both protocols and from their subsets, followed by analysis of gene expression using Affymetrix microarrays. Gene expression was compared between Ficoll and CPT samples for a given cell type.
Project description:High quality genetic material is an essential pre-requisite when analyzing gene expression using microarray technology. Peripheral blood mononuclear cells (PBMC) are frequently used for genomic analyses, but several factors can affect the integrity of nucleic acids prior to their extraction, including the methods of PBMC collection and isolation. In this study, we compared the Ficoll-Paque density gradient centrifugation and BD Vacutainer cell preparation tube (CPT) protocols to determine if either method offered a distinct advantage in preparation of PBMC-derived immune cell subsets for their use in gene expression analysis. We compared gene expression in PBMC and individual immune cell types from Ficoll and CPT isolation protocols using Affymetrix microarrays.
Project description:Human PB B cell subsets are functionally distinct and may derive from different developmental pathways, reflected by their differential gene expression profiles. We used microarrays to detail the global programme of gene expression underlying germinal center transition and functional propensities of individual subsets according to surface receptor and cell adhesion molecule expression. Human PB B cell subsets were sort-purified, RNA was extracted and processed. Data were vsn-normalized, corrected for batch-effects (ComBat) and analysed with GeneSpring GX7.