Project description:Gene expression profiles of CD34+CD38- stem cells and more differentiated CD34+CD38+ progenitor cells were compared. Comparison of expression profiles of hematopoietic stem cells from fetal liver, umbilical cord blood, bone marrow and mobilized pheripheral blood allowed us to identify a unique set of genes with conserved expression during ontogeny. Experiment Overall Design: CD34+CD38- en CD34+CD38+ cell populations were isolated by cell sorting from human Bone Marrow, mobilized peripheral blood, umbilical cord blood and fetal liver. Total RNA was isolated from each cell population followed by the synthesis of biotinylated cRNA. After fragmentation the biotinylated cRNA was hybridized to affymetrix U133A chips.
Project description:Gene expression profiles of CD34+CD38- stem cells and more differentiated CD34+CD38+ progenitor cells were compared. Comparison of expression profiles of hematopoietic stem cells from fetal liver, umbilical cord blood, bone marrow and mobilized pheripheral blood allowed us to identify a unique set of genes with conserved expression during ontogeny. Keywords: Cell type comparison
Project description:Comparison of the CD34+CD38- versus CD34+CD38+ fractions of human umbilical cord blood and comparison of the slow-dividing fraction versus the fast dividing fraction of the CD34+/CD38- population.
Project description:Evidence for the epigenetic regulation of hematopoietic stem cells (HSCs) is growing, but the genome-wide epigenetic signature of HSCs and its functional significance remain unclear. In this study, we used high-resolition (Agilent, CpG Island ChIP-on-chip) platforms for genome-wide comparison of CpG methylation in human CD34+ and CD34- cells. CD34+ and CD34- cells were purified from human umbilical cord blood. Genomic DNA obtained from CD34+ and CD34- cells was fragmented by either sonication or digestion with MseI. Recombinant methyl-binding domain (Genomictree, Daejeon, Korea) was incubated with sonicated genomic DNA in a binding reaction mixture. The methyl-enriched DNA fraction was purified using Qiaquick PCR purification kits (Qiagen, Hilden, Germany) and amplified using a whole genome amplification kit as recommended by Agilent. Amplified DNA products from CD34+ and CD34- cells were labeled with Cy3-dUTP and Cy5-dUTP, respectively, by random priming and hybridized onto Agilent human CpG island microarrays using conditions specified by the manufacturer (Agilent, Santa Clara, CA). After washing, the slides were scanned using an Agilent scanner and images were quantified using the Feature Extraction Software v. 9.3 (Agilent).
Project description:AML1-ETO expression in normal human umbilical cord blood CD34+ cells leads to long-term proliferation of an early self-renewing primitive progenitor cell with multilineage potential and stem cell ability, but these cells do not induce leukemia in immunodeficient mice. This comparative microarray study was initiated to determine the differences in the transcriptome of AML-ETO-expressing CD34+ cells after extended culture in vitro, using normal cord blood cells expanded for 6-8 weeks in vitro and subsequently purified for the CD34+ population as the control comparison. Keywords: Disease state analysis; comparison of changes in transcriptome due to long-term AML1-ETO expression in normal human hematopoietic CD34+ progenitor cells
Project description:Human cord blood CD34+ cells respond to the infection with LV by modifying the methylation profile of a large number of CpG sites in their genome. Different vector preparations vary in their capacity to trigger a response; some induce sporadic modifications only while others trigger a strong response that systematically includes de novo methylation of the same 4126 CpG-s corresponding to 1059 genes Illumina Infinium 450K arrays were performed according to the manufacturer's directions on bisulfite converted gDNA extracted from CD34+ cells from cord blood. CpG methylation profiling on Illumina Infinium 450K arrays was performed.
Project description:Genome wide DNA methylation profiling of 152 umbilical cord blood samples from CCCEH birth cohort. The Illumina Infinium 450k Human DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 450,000 CpGs in cord blood samples
