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Hematopoietic differentiation of human embryonic stem cells - timecourse and d7 CD34 and CD41 fractions

ABSTRACT: This study aimed to examine gene expression in human ES cells (the RUNX1C GFP reporter line) differentiated towrads hameatopoietic mesoderm in a defined serum free medium. At day 7 of differentiation, the cells were sorted into fractions based on CD34 and CD41 expression and the four fractions analysed by microarray.

ORGANISM(S): Homo sapiens

PROVIDER: GSE64876 | GEO | 2015/01/13

SECONDARY ACCESSION(S): PRJNA272402

REPOSITORIES: GEO

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Hematopoietic differentiation of human embryonic stem cells - timecourse and d7 CD34 and CD41 fractions

Project description:This study aimed to examine gene expression in human ES cells (the RUNX1C GFP reporter line) differentiated towrads hameatopoietic mesoderm in a defined serum free medium. At day 7 of differentiation, the cells were sorted into fractions based on CD34 and CD41 expression and the four fractions analysed by microarray. The total number of samples analysed was 13. Undifferentiated hESC (RUNX1C GFP/w, based on the HES3 cell line) plus samples from d1 to d8 of differentiation comprised one experiment (9 samples) and four flow sorted fractions from d7 differentiated cells (CD34-CD41-, CD34lo CD41-, CD34hi CD41- and CD34lo CD41lo) comprised the second experiment. The parent cell line was maintained on mouse feeder cells in KOSR containing medium supplemented with 10 ng/ml FGF2. Differentiation was performed as spin EBs in APEL medium (Ng et al Nature Protocols 2008). For the first 4 days, medium was supplemented with BMP4, VEGF, SCF and Activin. Medium was changed at d4 to fresh APEL medium supplemented with BMP4, VEGF, SCF, FGF2 and IGF2.

2015-01-13 | E-GEOD-64876 | biostudies-arrayexpress

Single cell RNA sequencing of CD41+ and CD41- CD34+ HPC in model of RUNX1 insufficiency

Project description:Single cell RNA sequencing of CD41+ and CD41- CD34+ HPC in model of RUNX1 insufficiency

| PRJNA699017 | ENA

Transcription profiling of human CD34- and + fractions from AML cells

Project description:CD34-positive and negative fractions were used for genomic DNA preparation, which was hybridized to HGU133 Plus 2.0 arrays. Experiment Overall Design: Samples with a suffix "-C" are CD34-negative controls, and those with a suffix "-L" are CD34-positive leukemic fractions. A total of 23 pairs were analyzed.

2008-06-13 | E-GEOD-4602 | biostudies-arrayexpress

Transcription profiling of human CD34+ fractions from AML for chromosome copy number analysis

Project description:CD34-positive fractions were used for mRNA preparation, which was hybridized to HGU 133 Plus 2.0 arrays. Experiment Overall Design: CD34-positive fractions of 22 AML patients were analyzed.

2008-06-13 | E-GEOD-4608 | biostudies-arrayexpress

Transcription profiling of CD34+CD38- vs CD34+CD38+ fractions of human umbilical cord blood and comparison of slow-dividing fraction vs the fast dividing fraction of CD34+/CD38- population

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.

2005-09-08 | E-EMBL-1 | biostudies-arrayexpress

Hematopoietic progenitors from differentiating RUNX1cGFP human embryonic stem cells

Project description:In order to identify cells expressing RUNX1c during hematopoietic differentiation of human embryonic stem cells (hESCs), we targeted GFP downstream of the RUNX1 distal promoter. GFP was observed from 10-25 days of embryoid body differentiation and accurately mirrored expression of the endogenous RUNX1c isoform. GFP was restricted to CD45+ hematopoietic cells and GFP+CD34+ cells were highly enriched for progenitor cells. Appearance of the first wave of hematopoietic blast colony forming cells (Bl-CFC) in d3-4 embryoid bodies, antedated the expression of RUNX1c. These results confirm a role for RUNX1c in marking a second wave of hematopoietic progenitor cells in differentiating hESCs. The purpose of the cord blood samples was to perform comparison of them to the gene expression profiles of the Runx fractions - as the cord blood represents a heterogeneous mix of hematopoietic stem cells and multipotent progenitors cells.

2012-02-12 | E-MEXP-3356 | biostudies-arrayexpress

Differentiation of human embryonic stem cells to HOXA+ hemogenic vasculature that resembles the aorta-gonad-mesonephros

Project description:The production of definitive haematopoietic stem/progenitor cells from human pluripotent stem cells (hPSCs) remains a significant challenge. Using reporter lines to track the endothelial (SOX17) to haematopoietic (RUNX1C) transition, we found that hPSC differentiated in growth factor supplemented serum free medium generated RUNX1C+CD34+ clonogenic cells that homed to the bone marrow, but did not engraft. Compared to repopulation-competent cord blood CD34+ cells, RUNX1C+CD34+ progenitors lacked HOXA gene expression, indicating incorrect mesoderm patterning. This deficiency was ameliorated by a timed pulse of WNT activation combined with ACTIVIN antagonism. Significantly, these HOXA+ cultures now formed complex SOX17+ vessels that produced fetal liver-like haematopoietic cells, similar to the human aorta-gonad-mesonephros (AGM). Comparison of transcriptional profiles of these nascent haematopoietic stem/progenitors with cells isolated from human AGM confirmed significant similarities, consistent with the assignment of our in vitro generated cells to the definitive human haematopoietic lineage. Our findings argue that HOXA codes established early in differentiation predict cellular potential and provide correct cell patterning for the specification of definitive haematopoietic lineages from hPSCs.

2016-10-17 | GSE81080 | GEO

Differentiation of human embryonic stem cells to HOXA+ hemogenic vasculature that resembles the aorta-gonad-mesonephros

2016-10-17 | GSE81074 | GEO

Clostridioides difficile CD41

Project description:Clostridioides difficile CD41 Genome sequencing and assembly

| PRJNA85701 | ENA

Gene expression analyses of hematopoietic stem cell (HSC) subsets in wildtype or CD41-KO mice

Project description:The hematopoietic stem cell (HSC) compartment is heterogeneous, yet our understanding of the identities of different HSC subtypes is limited. Here we show that platelet integrin CD41 (αIIb), currently thought to only transiently mark fetal HSCs, is expressed on an adult HSC subtype that accumulates with age. CD41+ HSCs were largely quiescent and exhibited myeloerythroid and megakaryocyte gene priming, governed by Gata1, whereas CD41- HSCs were more proliferative and exhibited lymphoid gene priming. When isolated without the use of blocking antibodies, CD41+ HSCs possessed long-term repopulation capacity upon serial transplantations and showed a marked myeloid-bias compared to CD41-HSCs, which yielded a more lymphoid-biased progeny. CD41-knockout mice displayed multilineage hematopoietic defects coupled with decreased quiescence and survival of HSCs, suggesting that CD41 is functionally relevant for HSC maintenance and hematopoietic homeostasis. Transplantation experiments indicated that CD41-KO associated defects are long-term transplantable and HSC-derived, and in part mediated through the loss of platelet mass leading to decreases in HSC exposure to important platelet released cytokines, such as TGFβ1. In summary, our data provide a novel marker to identify a myeloid-biased HSC subtype that becomes prevalent with age, and highlights the dogma of HSC regulation by their progeny.

2013-04-11 | GSE45561 | GEO

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