Timecourse of in vitro erythroid differentiation of human adult-derived peripheral blood CD34+ cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE4655 | GEO | 2006/11/09
SECONDARY ACCESSION(S): PRJNA95299
REPOSITORIES: GEO
Similar Datasets
Project description:The aim of the study is to develop a scalable continuous agitation suspension culture bioprocess for differentiation of hiPSCs towards erythroid cells. Using process optimization, we developed the entire process completely in agitation suspension culture condition starting from hiPSC expansion through to hematopoietic induction and erythroid differentiation. Transcriptome profiling was carried out to facilitate comparisons between erythroid cells differentiated from hiPSCs using this bioprocess, and erythroid cells differentiated from cord blood and adult CD34+ cells.
2021-03-03 | GSE137917 | GEO
Project description:The Affymetrix Human Gene 2.0 ST array was used to measure differential expression of RNA isolated from normal and Diamond Blackfan anemia (DBA) erythroid progenitors after ex vivo expansion of circulating, peripheral blood derived hematopoietic stem cells under erythroid growth conditions. The gene-level probe summaries reported in this series were computed using RMA as implemented in the Bioconductor package Oligo v1.36.1. Diamond Blackfan anemia (DBA) is a congenital bone marrow failure syndrome characterized by erythroid aplasia, usually without perturbation of other hematopoietic lineages. Approximately 65% of DBA patients with autosomal dominant inheritance have heterozygous mutations or deletions in ribosomal protein (RP) genes while <1% of patients with X-linked inheritance have been identified with mutations in the transcription factor, GATA1. Erythroid cells from patients with DBA have not been well characterized and the mechanisms underlying the erythroid specific effects of either RP or GATA1 associated DBA remain unclear. We have developed an in vitro culture system to expand peripheral blood CD34+ progenitor cells from patients with DBA and differentiate them into erythroid cells. Cells from patients with RP or GATA1 mutations showed decreased proliferation and delayed erythroid differentiation compared to controls. RNA transcript analyses of erythroid cells from controls and patients with RP or GATA1 mutations showed distinctive differences, with upregulation of heme biosynthesis genes prominently in RP-mediated DBA and failure to upregulate components of the translational apparatus in GATA1-mediated DBA. Our data show that dysregulation of translational function is a common feature of DBA caused by both RP and GATA1 mutations.
2017-04-04 | GSE89540 | GEO
Project description:computational analysis of transcriptomic information during erythroid development Experiment Overall Design: mRNA expression profiling was used to characterize developing erythroid cells at 6 time points during differentiation.
2007-11-02 | E-GEOD-4655 | biostudies-arrayexpress
Project description:Analysis of mobilized peripheral blood CD34+ cells from a healthy volunteer under erythroid differentiation conditions with and without stimulation to the BMP or Wnt signaling pathways. For erythroid differentiation, expanded CD34+ cells were placed in Stemspan SFEM medium supplemented with 2% pen/strep, 20ng/ml SCF, 1U/ml Epo, 5ng/ml IL3, 2uM dexamethasone, and 1uM beta-estradiol. Arrays were performed 2 hours after addition of cytokines. For signaling pathway stimulation, cells were exposed to 0.5uM BIO (a GSK3 inhibitor) for Wnt pathway activation, 25ng/ml rhBMP4 for BMP pathway activation, or vehicle control for 2 hours. Three biological replicates were performed per treatment group. We used microarrays to detail the global program of gene expression changes after Wnt or BMP pathway stimulation in human CD34+ hematopoietic progenitors under erythroid differentiation conditions.
2011-10-07 | GSE26351 | GEO
Project description:Analysis of mobilized peripheral blood CD34+ cells from a healthy volunteer under erythroid differentiation conditions with and without stimulation to the BMP or Wnt signaling pathways. For erythroid differentiation, expanded CD34+ cells were placed in Stemspan SFEM medium supplemented with 2% pen/strep, 20ng/ml SCF, 1U/ml Epo, 5ng/ml IL3, 2uM dexamethasone, and 1uM beta-estradiol. Arrays were performed 2 hours after addition of cytokines. For signaling pathway stimulation, cells were exposed to 0.5uM BIO (a GSK3 inhibitor) for Wnt pathway activation, 25ng/ml rhBMP4 for BMP pathway activation, or vehicle control for 2 hours. Three biological replicates were performed per treatment group. We used microarrays to detail the global program of gene expression changes after Wnt or BMP pathway stimulation in human CD34+ hematopoietic progenitors under erythroid differentiation conditions. To investigate the changes to gene expression in human CD34+ hematopoietic progenitors following stimulation of the Wnt or BMP pathways during the early stages of erythroid differentiation. Three biological replicates were performed per treatment group.
2011-10-06 | E-GEOD-26351 | biostudies-arrayexpress
Project description:Ex vivo manufactured red blood cells (RBC) generated from immortalized erythroid cell lines which can semi-infinitely grow are expected to become a significant alternative in future transfusion therapies. To establish those cell lines, ectopic expression of human papilloma virus (HPV) E6/E7 gene has successfully been employed. In order to induce differentiation and maturation of the immortalized cell lines, terminating the HPV-E6/E7 expression through a gene induction system has been believed to be essential. Here we report that erythroid cell lines established from human bone marrow using simple overexpression of HPV-E6/E7 is capable of normal erythroid differentiation without turning the gene expression off. Newly established cell lines, Erythroid Lines from Lund University (ELLU), are able to differentiate towards mature cells including enucleated reticulocytes upon changing the culture condition but without terminating HPV-E6/E7 gene expression. ELLU cells are heterogeneous, and unexpectedly, clones expressing adult hemoglobin rapidly differentiate but produced cells are fragile while other clones with fetal hemoglobin start expressing adult hemoglobin upon differentiation and give rise to more mature cells. Our findings propose an alternative method to establish immortalized human erythroid cell lines and describe novel cellular characteristics for desired functionally competent clones.
2022-01-31 | GSE184083 | GEO
Project description:Bipotent megakaryocyte/erythroid progenitors (MEPs) give rise to progeny limited to the megakaryocyte (Mk) and erythroid (E) lineages. We developed a novel dual-detection functional in vitro colony-forming unit (CFU) assay for single cells that differentiates down both the Mk and E lineages (CFU-Mk/E), which allowed development and validation of a novel purification strategy for the identification and quantitation of primary functional human MEPs from granulocyte colony-stimulating factor-mobilized peripheral blood and bone marrow. Applying this assay to fluorescence-activated cell sorter-sorted cell populations, we found that the Lin(-)CD34(+)CD38(mid)CD45RA(-)FLT3(-)MPL(+)CD36(-)CD41(-) population is much more highly enriched for bipotent MEPs than any previously reported subpopulations. We also developed purification strategies for primary human lineage-committed Mk and E progenitors identified as CFU-Mk and burst forming unit-E. Comparative expression analyses in MEP, MkP, and ErP populations revealed differential expression of MYB We tested whether alterations in MYB concentration affect the Mk-E fate decision at the single cell level in MEPs and found that short hairpin RNA-mediated MYB knockdown promoted commitment of MEPs to the Mk lineage, further defining its role in MEP lineage fate. There are numerous applications for these novel enrichment strategies, including facilitating mechanistic studies of MEP lineage commitment, improving approaches for in vitro expansion of Mk and E cells, and developing improved therapies for benign and malignant hematologic disease.
| S-EPMC4990855 | biostudies-other
Project description:Transcriptome analysis of human erythroid cells differentiated from iPSCs, cord blood and adult peripheral blood
| PRJNA573880 | ENA
Project description:Advances in sequencing-based genomic profiling present a new challenge of explaining how changes in DNA/RNA are translated into proteins linking genotypes to phenotypes. The developing erythroid cells require highly coordinated gene expression and metabolism, and serve as a unique model in dissecting regulatory events in development and disease. Here we compare the proteomic and transcriptomic changes in human hematopoietic stem/progenitor cells and lineage-committed erythroid progenitors, and uncover pathways related to mitochondrial biogenesis enhanced through post-transcriptional regulation. Two principal mitochondrial factors TFAM and PHB2 are tightly regulated at the protein level and indispensable for mitochondria and erythropoiesis. mTORC1 signaling is progressively enhanced to promote translation of mitochondrial proteins during erythroid specification. Genetic and pharmacological perturbation of mTORC1 or mitochondria impairs erythropoiesis. Our studies suggest a new mechanism for regulation of mitochondrial biogenesis through mTORC1-mediated protein translation, and may have direct relevance to the hematological defects associated with mitochondrial diseases and aging. Transcriptional profiling in human primary fetal and adult CD34+ hematopoietic stem/progenitor cells (HSPCs) erythroid progenitor cells (ProEs) by RNA-seq analysis.
2017-05-15 | E-GEOD-74053 | biostudies-arrayexpress
Project description:We provide the first full account of transcriptional changes during terminal erythroid differentiation for cells grown under conditions of stress erythropoiesis. Samples were generated from PBMC's in three-stage culture system in Cellquin, an IMDB based culture medium optimized for erythroid outgrowth. The first two stages rely on presence of EPO Stem Cell Factor and dexamethasone (mimmicking stress erythropoiesis), followed by a terminal differentiation stage in presence of EPO and Holotransferrin. This culture set up allows for generation of large number of cultured Red Blood Cells that ar functionally and morphologically similar to ex-vivo red blood cells.
2019-11-25 | GSE124363 | GEO