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Expression Profiling of Primary Human Fetal and Adult Hematopoietic Stem/Progenitor Cells (HSPCs) and Differentiating Proerythroblasts (ProEs)

ABSTRACT: To facilitate comparative genomic analyses of human fetal and adult cells undergoing erythropoiesis, we employed a serum-free two-phase liquid culture system to expand and differentiate primary human CD34+ hematopoietic stem/progenitor cells (HSPCs) ex vivo. In this experimental context, highly enriched populations of stage-matched, differentiating, primary proerythroblasts (ProEs) were generated. We selected four time points (day 0, CD34+ HSPCs; day 3, 5, and 7, differentiating ProEs) that represented similar stages differentiation for gene expression profiling using microarrays.

ORGANISM(S): Homo sapiens

PROVIDER: GSE36984 | GEO | 2012/10/05

SECONDARY ACCESSION(S): PRJNA157467

REPOSITORIES: GEO

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Expression Profiling of Primary Human Fetal and Adult Hematopoietic Stem/Progenitor Cells (HSPCs) and Differentiating Proerythroblasts (ProEs)

Project description:To facilitate comparative genomic analyses of human fetal and adult cells undergoing erythropoiesis, we employed a serum-free two-phase liquid culture system to expand and differentiate primary human CD34+ hematopoietic stem/progenitor cells (HSPCs) ex vivo. In this experimental context, highly enriched populations of stage-matched, differentiating, primary proerythroblasts (ProEs) were generated. We selected four time points (day 0, CD34+ HSPCs; day 3, 5, and 7, differentiating ProEs) that represented similar stages differentiation for gene expression profiling using microarrays. Primary maturing fetal or adult erythroblasts were generated ex vivo from CD34+ hematopoietic stem/progenitor cells (HSPCs) using a serum-free two-phase liquid culture system. Total RNA from primary fetal and adult HSPCs (day 0) and differentiating proerythroblasts (ProEs; day 3, 5, and 7) were extracted and used to hybridize to Affymetrix expression arrays using the HG-U133 Plus 2.0 platform.

2012-10-05 | E-GEOD-36984 | biostudies-arrayexpress

Expression Profiling of Primary Human Proerythroblasts (ProEs) After IRF2, IRF6, and MYB shRNA Knockdown

Project description:IRF2, IRF6, and MYB are candidate regulators of human erythropoiesis. We here examine primary CD34+ hematopoietic stem/progenitor cells (HSPCs)-derived erythroid progenitors with control, IRF2, IRF6, or MYB shRNA lentiviral transduction prior to differentiation. Gene expression microarray profiling datasets for MYB shRNA and control shRNA were obtained from Gene Expression Omnibus (GEO) under accession number GSE25678. The data were analyzed together with the datasets obtained in this study. Primary maturing adult erythroblasts were generated ex vivo from CD34+ hematopoietic stem/progenitor cells (HSPCs) using a serum-free two-phase liquid culture system. CD34+ HSPCs were transduced with lentiviruses containing shRNAs against IRF2 or IRF6 gene, selected and differentiated to proerythroblasts (ProEs). Cells were harvested at day 5 of differentiated and total RNA were extracted. This was used to hybridize to Affymetrix expression arrays using the HG-U133 Plus 2.0 platform.

2012-10-05 | E-GEOD-36988 | biostudies-arrayexpress

Comparative profiling of human fetal and adult erythropoiesis

Project description:This SuperSeries is composed of the following subset Series: GSE36984: Expression Profiling of Primary Human Fetal and Adult Hematopoietic Stem/Progenitor Cells (HSPCs) and Differentiating Proerythroblasts (ProEs) GSE36985: Comparative profiling of chromatin state maps and transcription factor occupancy during human fetal and adult erythropoiesis GSE36988: Expression Profiling of Primary Human Proerythroblasts (ProEs) After IRF2, IRF6, and MYB shRNA Knockdown Refer to individual Series

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RNA-seq transcriptional profiling in human primary fetal and adult CD34+ hematopoietic stem/progenitor cells (HSPCs) erythroid progenitor cells (ProEs)

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

Homo sapiens

Project description:Expression Profiling of Primary Human Fetal and Adult Hematopoietic Stem/Progenitor Cells (HSPCs) and Differentiating Proerythroblasts (ProEs)

| PRJNA157467 | ENA

RNA-seq transcriptonal profiling in human primary adult erythroid progenitor cells (ProEs) after shRNA-mediated depletion of TFAM and PHB2 expression

Project description:The developing erythroid cells require highly coordinated gene expression and metabolism. By comparing the proteomic and transcriptomic changes in human hematopoietic stem/progenitor cells (HSPCs) and lineage-committed erythroid progenitors (ProEs), 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. To determine the role of TFAM and PHB2 in mitochondrial function during erythroid development, we employed shRNA-mediated depeltion of TFAM and PHB2 expression in differentiating erythroid cells, and performed RNA-seq transcriptional profiling analysis.

2017-05-15 | GSE86910 | GEO

Murine Proerythroblasts (ProEs): WT vs. Bmi1-/-

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2014-11-18 | GSE63413 | GEO

DARTseq identifies m6A targets in human CD34+ HSPCs

Project description:The goal of this study is to identify m6A targets in human CD34+ HSPCs with or without STM2457 treatment using DARTseq. Purified human CD34+ HSPC cells were transduced with APOBEC-YTH or empty vector control (MIG). 24hr post infection, CD34+ cells were treated with STM2457 at a concentration of 20µM for 2 days. GFP+ cells were sorted the next day. 3 replicates were performed.

2023-12-07 | GSE231861 | GEO

Murine Proerythroblasts (ProEs): WT vs. Bmi1-/-

Project description:Transcriptional profiling of ProEs purified from wild type and Bmi1-/- mice. Two-condition experiment, WT vs. Bmi1-/- ProEs. Biological replicates: 3 WT replicates, 3 Bmi1-/- replicates.

2014-11-18 | E-GEOD-63413 | biostudies-arrayexpress

Next Generation Sequencing Analysis of human hematopoietic stem/progenitor cells (HSPCs) differentiated from pluripotent stem cells

Project description:Purpose: global gene expression profiling of H1 and iPSC derived HSPCs. Methods: SMART-seq2 amplified polyA RNA from differentiation day 8 CD31+CD34+CD43+ and CD31+CD34+CD43- cells, and day 5 CD31+CD34+ cells cultured in medium with or without insulin supplement Results: Modulation of key cell signaling pathways lead to highly efficient HSPC differentiation from human pluripotent stem cells. Conclusions: Highly efficient HSPC differentiation from human pluripotent stem cells in a monolayer, chemically defined culture system.

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