Project description:Expression data from Notch-activated CD34+CD45RA-Lin- hematopoietic progenitor cells (HPCs) transduced with nuclear-trapped AF1q/MLLT11 (A2M)
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 CD34+CD45RA-Lin- and CD34+CD45RA+Lin- HPCs were FACS-sorted, exposed to A2M or control vectors, sorted based on GFP expression, and subjected to global gene expression analysis 72 hrs later.
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 CD34+CD45RA-Lin- HPCs were FACS-sorted, exposed to A2M or control vectors, sorted again based on GFP expression, and cultured for 72 hrs with graded doses of plastic-immobilized Notch ligand Delta1ext-IgG
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: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:Group 3 innate lymphoid cells (ILC3) are defined by the expression of RORM-NM-3t, which is selectively required for their development. The lineage-specified progenitor cells of human ILC3 and their developmental site after birth remain undefined. Here we identified a novel population of human CD34+ hematopoietic progenitor cells (HPC) expressing RORM-NM-3t and sharing with ILC3 a distinct transcriptional signature. RORM-NM-3t+ CD34+ HPC were located in tonsils and intestinal lamina propria (LP) and selectively differentiated towards ILC3. Conversely, RORM-NM-3t- CD34+ HPC displayed commitment potential for both ILC3 and NK cells and the differentiation fate towards these two cell lineages was determined by cytokine and aryl hydrocarbon receptor (AhR) signaling. Thus, we propose that RORM-NM-3t+ CD34+ cells represent human lineage-specified progenitors of IL-22+ ILC3 and that tonsils as well as intestinal LP might be preferential sites of their differentiation. ILC3, NK cells and the CD34+ HPC subsets were sorted from tonsils of 6 distinct donors to purity above 95%. cRNA of the sorted cell populations was hybridized to an Agilent Whole Human Genome Oligo Microarrays (8x60K v2, Design ID 039494)
Project description:Group 3 innate lymphoid cells (ILC3) are defined by the expression of RORγt, which is selectively required for their development. The lineage-specified progenitor cells of human ILC3 and their developmental site after birth remain undefined. Here we identified a novel population of human CD34+ hematopoietic progenitor cells (HPC) expressing RORγt and sharing with ILC3 a distinct transcriptional signature. RORγt+ CD34+ HPC were located in tonsils and intestinal lamina propria (LP) and selectively differentiated towards ILC3. Conversely, RORγt- CD34+ HPC displayed commitment potential for both ILC3 and NK cells and the differentiation fate towards these two cell lineages was determined by cytokine and aryl hydrocarbon receptor (AhR) signaling. Thus, we propose that RORγt+ CD34+ cells represent human lineage-specified progenitors of IL-22+ ILC3 and that tonsils as well as intestinal LP might be preferential sites of their differentiation.
Project description:CD34+ HPC cells were retrovirally transduced with either control PINCO-GFP or PINCO Gamma-Catenin-GFP and analysed for Affymetrix mRNA expression. Gamma-Catenin and control matched transduced cells were also cultured until day 6 and were fractionated into the main progenitor groups for Affymetrix microarray analysis as previously described (tonks 2007). Briefly, a depletion strategy using the Miltenyi Biotec MiniMACS system was used for this purpose where, myeloblasts (CD15+) were enriched from mixed lineage transduced cells by serial depletion of monoblasts (CD14hi) and erythroblasts (CD36+) using immunomagnetic beads directed against those antigens.
Project description:CD34+ hematopoietic stem/progenitor cells were isolated from human cord blood and amplified in vitro for 10-14 days in serum-free medium with specific cytokines (Ju et al., Eur. J. Cell Biol. 82, 75-86, 2003; Hacker et al., Nat. Immunol. 4, 380-386, 2003). Cells were then treated with TGF-beta1 for various periods of time (2, 4, 16 hours) and RNA was prepared and subjected to microarray analysis. Experiment Overall Design: CD34+ hematopoietic stem/progenitor cells (HPC) were amplified in vitro and treated with TGF-beta1 (10 ng/ml) for 2, 4 and 16 hours. Experiment Overall Design: HPC untreated Experiment Overall Design: HPC + TGF-beta1 for 2 hours Experiment Overall Design: HPC + TGF-beta1 for 4 hours Experiment Overall Design: HPC + TGF-beta1 for 16 hours
