Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA272402 | ENA |
REPOSITORIES: ENA
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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
Project description:Human CD34+ progenitors can be in vitro differentiated into proplatelet-producing megakaryocytes (MKs) within 17 days. During this course, 4 cell populations emerge, phenotypically defined as CD34+CD41+ at day 7 (D7) and CD34+CD41+CD9- at D10 and D14 —qualified as “productive” because they can differentiate into proplatelet-forming cells during the D14-D17 period— and CD34-CD41+ or CD34+CD41+CD9+ at day 10 —qualified as “unproductive”, because unable to form proplatelets later. The productive pathway is boosted by the addition of SR1 at D0 and D7. To clarify the features of the productive and unproductive pathways, as well as the effect of SR1, the transcriptomes of the cell populations present at D0, 7, 10 and 14, generated in the presence of the absence of SR1 were determined by RNA-Seq.
2021-06-30 | GSE167866 | GEO
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.
2015-01-13 | GSE64876 | GEO
Project description:CD34 positive hematopoietic stem cells were differentiated into erythroid lineage. Next generation sequencing (NGS) of 5hmC affinity pulldown and RNAseq were performed in four time point of different stages of erythroid differentiation. 4 RNA-Seq Samples (d0, d3, d7 and d10); 4 affinity-pulldown (d0, d3, d7 and d10), and 4 input samples (d0, d3, d7 and d10).
2014-03-28 | E-GEOD-40243 | biostudies-arrayexpress
Project description:Cord blood CD34+ hematopoietic precursors (3 donors) were control or HES6 shRNA (HES6 knockdown) transduced and cultured for four days in media containing SCF, TPO and EPO to drive differentiation towards megakaryocytes and erythroid cells. After four days four subpopulations were sorted for RNA isolation to research the effect of HES6 knockdown on gene expression. We sorted megakaryocytes (CD41+) and early (CD71+ CD235-) and late (CD71+ CD235+) erythroblasts (CD41-) and CD34- precursor cells (CD41- CD71- CD235- CD34-).
2024-11-13 | GSE229196 | GEO
Project description:RNA-seq of endothelial (Endo), Pre-hematopoietic progenitor cells (Pre-HPCs) and hematopoietic progenitors (HP) derived from the in-vitro hematopoietic differentiation of mouse Embryonic Stem Cells (mESCs) harboring a biallelic inactivating deletion of the endogenous Tal1 gene, and a construct for the dox-induced expression of the hematopoietic genes Tal1, Lyl1 and Lmo2 (i3TFs Tal1Δ/Δ mESCs). i: inducible. TFs: transcription factors. Cells were treated with dox at one (Dox Unt) or two (Dox Dox) time-points during hematopoietic differentiation to inducibly express the 3TFs. Sequenced cell populations were purified by FACS sorting based on the cell-surface expression of the endothelial marker VE-CADHERIN and the hematopoietic marker CD41. Endo: VE-CADHERIN+CD41- Pre-HPCs: VE-CADHERIN+CD41+ HP: VE-CADHERIN-CD41+
2023-05-15 | E-MTAB-12973 | biostudies-arrayexpress
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
Project description:Ex vivo differentiation of megakaryocytes (Mk) was carried out using human cord blood (CB) CD34+ cells under the stimulation of recombinant human interleukin-3, stem cell factor, and thrombopoietin for 7 days, followed by thrombopoietin treatment alone for further 3 days. Total cellular RNA was extracted from Day-0 CD34+, Day-10 CD41+ and CD41- cells, respectively. Microarray was performed and the data was analyzed using the GeneChip Operating Software, Spotfire DecisionSite Software and Genomatix Application Software.
2009-03-25 | E-MEXP-1546 | biostudies-arrayexpress
Project description:Epigenetic memory in induced pluripotent stem cells (iPSCs), with regards to their somatic cell type of origin, might lead to variations in their differentiation capacities. In this context, iPSCs from human CD34+ hematopoietic stem cells (HSCs) might be more suitable for hematopoietic differentiation than commonly used fibroblast-derived iPSCs. To investigate the influence of an epigenetic memory on the ex vivo expansion of iPSCs into erythroid cells, we compared iPSCs from human neural stem cells (NSCs) and human cord blood-derived CD34+ HSCs and evaluated their potential for differentiation into hematopoietic progenitor and mature red blood cells (RBCs). Although genome-wide DNA methylation profiling at all promoter regions demonstrates an epigenetic memory of iPSCs with regards to their somatic cell type of origin, we found a similar hematopoietic induction potential and erythroid differentiation pattern. All human iPSC lines showed terminal maturation into normoblasts and enucleated RBCs, producing predominantly fetal hemoglobin. Differences were only observed in the growth rate of erythroid cells, which was slightly higher in the CD34+ HSC-derived iPSCs. More detailed methylation analysis of the hematopoietic and erythrocyte promoters identified similar CpG methylation levels in the CD34+ iPSCs and NSC iPSCs, which confirms their comparable erythroid differentiation potential. To investigate the influence of an epigenetic memory on the ex vivo expansion of iPSCs into erythroid cells, we compared iPSCs from human neural stem cells (NSCs) and human cord blood-derived CD34+ HSCs and evaluated their potential for differentiation into hematopoietic progenitor and mature red blood cells (RBCs). RNA samples for microarray analysis were prepared using RNeasy columns (Qiagen, Germany) with on-column DNA digestion. 300ng of total RNA per sample was used as the input in the linear amplification protocol (Ambion), which involved the synthesis of T7-linked double-stranded cDNAs and 12hrs of in vitro transcription incorporating the biotin-labeled nucleotides. Purified and labeled cRNA was then hybridized for 18hrs onto HumanHT-12 v4 expression BeadChips (Illumina, USA) following the manufacturer's instructions. After the recommended washing, the chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using the iScan reader (Illumina) and the accompanying software. The samples were exclusively hybridized as biological replicates. 8 samples were analyzed: CD34 1, Human CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) population 1, 1 replicate CD34 2, Human CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) population 2, 1 replicate CD34 OSiPS 1, Human Human two factors (POU5F1, SOX2) induced Pluripotent Cell (iPSC) reprogrammed from CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) induced Pluripotent Cell (iPSC) population 1, 1 replicate CD34 OSKMiPS 1, Human Human four factors (POU5F1, SOX2, KLF4, CMYC) induced Pluripotent Cell (iPSC) reprogrammed from CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) induced Pluripotent Cell (iPSC) population 1, 1 replicate CD34 OSiPS 2, Human Human two factors (POU5F1, SOX2) induced Pluripotent Cell (iPSC) reprogrammed from CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) induced Pluripotent Cell (iPSC) population 2, 1 replicate CD34 OSKMiPS 2, Human Human four factors (POU5F1, SOX2, KLF4, CMYC) induced Pluripotent Cell (iPSC) reprogrammed from CD34+ Cord blood CD34+ Hematopoyetic Stem Cell(HSC) induced Pluripotent Cell (iPSC) population 2, 1 replicate H1, Human H1 embryonic stem cell (ESC), 1 replicate H9, Human H9 embryonic stem cell (ESC), 1 replicate
2015-02-20 | E-GEOD-55109 | biostudies-arrayexpress
Project description:Ex vivo production of induced megakaryocytes (MKs) and platelets from stem cells is an alternative approach for supplying transfusible platelets. However, it is still difficult to generate large numbers of MKs and platelets from cord blood hematopoietic stem cells. To optimize the differentiation efficiency of megakaryocytic cells from hematopoietic stem and progenitor cells (HSPCs), we first employed a platelet factor 4 (PF4)-promoter reporter and high-throughput screening strategy to screen for small molecules. We found that a small molecule, Ricolinostat, efficiently promoted the generation of CD34+CD41+ megakaryocyte progenitor cells (MkPs) and CD41+CD61+ MKs from cord blood HSPCs. Ricolinostat showed the capacity to enhance the cell fate commitment of MkPs from HSPCs and promoted the proliferation of CD34+CD41+ MkPs. Ricolinostat significantly increased the gene expression levels of key MK transcriptional factors, including HOXC6 and PCGF2. Notably, these megakaryocytic cells generated from Ricolinostat-induced HSPCs could differentiate into mature MKs and platelets. Additionally, RNA sequencing data suggested that Ricolinostat might enhance MkP fate mainly by inhibiting the secretion of IL-8 and decreasing the expression of IL-8 receptor CXCR2. Our study will help in the development of manufacturing protocols for large-scale generation of induced MKs and platelets for clinical application.
2022-02-09 | GSE182181 | GEO