Project description:This SuperSeries is composed of the following subset Series: GSE40829: Expression profiles of lineage-depleted (Lin-) cell and mono-nucleated cell (MNC) samples derived from human umbilical cord blood GSE40830: Expression analysis of uncultured and culture-derived colony forming unit-monocytes and megakaryocytes Refer to individual Series
Project description:Despite the importance of inter-cellular communication networks in regulating stem cell fate decisions, very little is known about the topology, dynamics, or functional significance. Using human blood stem cell cultures as an experimental paradigm, we present a novel bioinformatic approach to integrate genome-scale molecular profiles (transcriptome and secretome) and publicly available databases to reconstruct soluble factor-mediated inter-cellular signalling networks regulating blood stem cell fate decisions. Umbilical cord blood UCB cells were lineage depleted via immuno-magnetic column and placed in serum free media, supplemented with SCF, FL, and TPO. At culture day 4, cells were either subject to Lin- cell re-selection + media exchange (S/E) or not (NS/NE) and cultured for an additional 4 days. At day-8, S/E cell sample was subject to another S/E procedure and cultured for an additional 4 days. At day-0, 4, 8, and 12, cells were separated into Lin- and Lin+ for independent profiling using Affymetrix HU133 v2.0 arrays.
Project description:Stem/progenitor cells demonstrate neuro-regenerative potential that is dependent upon their humoral activity by producing various trophic factors that regulate cell migration, growth, and differentiation. To better characterise the human umbilical cord blood-derived stem/progenitor cells, we analyzed the global gene expression pattern in the lineage-negative, CD34+, CD133+ cells and unseparated nucleated cells. We were interested in biological processes concerning biosynthetic processes, cytokine production, secretion by cells, chemotaxis, migration, and proliferative capacities.
Project description:Lineage-depleted (Lin-) cell and mono-nucleated cell (MNC), and uncultured and culture-derived colony forming unit-monocytes and megakaryocytes
Project description:We used microarrays to examine the impact of AF1q/MLLT11 on the gene expression profile of CD34+CD45RA-Lin- and CD34+CD45RA+Lin- HPCs isolated from umbilical cord blood CD34+CD45RA-Lin- and CD34+CD45RA+Lin- HPCs correspond respectively to early multipotent and lympho-granulo-macrophagic precursors. A2M is a nuclear mutant-derivative of AF1q/MLLT11
Project description:Genome-wide DNA methylation (DNAm) studies have been extremely useful to understand hematopoiesis in humans. For cell-specific epigenetic studies, fluorescence-activated cell sorting (FACS) is the gold standard technique to isolate homogeneous cell populations of interest. However, in cord blood, the DNAm signature of isolated hematopoietic cells may be significantly altered by heterotopic interactions with nucleated red blood cells (nRBCs) that go undetected during conventional cell sorting. Using the Illumina 450K array, genome-wide DNAm profiles of the following cell types were obtained: (1) T lymphocytes, monocytes, and nRBCs isolated using a “standard” strategy lacking exclusion for erythroid lineage markers; and (2) CD4 and CD8 T lymphocytes, B lymphocytes, Natural Killer (NK) cells, granulocytes, monocytes, and nRBCs isolated using a “stringent” strategy formally excluding erythroid lineage-specific markers. DNAm profiles of cord blood cells isolated by the standard sorting strategy showed significant heterotopic cross-contamination between cell populations, whereas the cord blood cells sorted by the stringent strategy displayed DNAm profiles more consistent with their expected hematopoietic lineage relationships.
Project description:We used microarrays to examine the impact of AF1q/MLLT11 on the gene expression profile of Notch-activated CD34+CD45RA-Lin- HPCs isolated from umbilical cord blood CD34+CD45RA-Lin- HPCs correspond to early multipotent progenitors. A2M is a nuclear mutant derivative of AF1q/MLLT11
Project description:Expansion for hematopoietic cells from umbilical cord blood is a strategy for use this cell source in clinic transplants, however, it is important to know about the genomic changes that can occur in expanded cells. In order to detect global expression profiles changes in hematopoietic stem and progenitors cells generated in vitro, we analyzed hematopoietics populations obtained by FACS in fresh from umbilical cord blood. HSC (fHSC) was defined as CD34+ CD38- CD71- CD45RA- Lin- and were cocultured with stromal cell line OP-9 plus FL, SCF, IL3, IL6, TPO, GMCSF and G-CSF by 7 days, after time we repurified HSC population by FACS using same immunophenotype (ivHSC). In other hand, fresh erythroid progenitors cells (fEPC) were identified as CD34+CD38+CD71+CD45RA- Lin- and fresh myeloid progenitors cells (fMPC) were identified as CD34+CD38+CD71-CD45RA+Lin-. In vitro progenitors cells (ivEPC and ivMPC) were obtained by culturing fHSC in Stemspan serum-free media plus SCF, TPO, IL6, FL and IL3 by 10 days, after time cells were repurified by FACS using same immunophenotype for fresh progenitors. In vitro generated cells were compared with their corresponding fresh population cells.
