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Single cell RNA-sequencing (scRNA-seq) of the human hematopoietic stem cell compartment (CD34+CD38-CD45RA-)

ABSTRACT: To identify a trajectory of human hematopoietic stem cell (HSC) activation single cell gene expression profiling of RNA from human HSC was performed by flow cytometry-based single cell deposition of CD34+CD38Ã¢Â€Â”CD45RAÃ¢Â€Â”cells from G-CSF-mobilized peripheral blood from 3 individual donors and 1 bone marrow donor in concert with single cell index-sorting (signals indexed: CD34, CD38, CD45RA, CD90, CD49f) and followed by SMARTseq2 based protocol. We show that INKA1 and CDK6/PAK4 expressing single cells segregate across alternative states of quiescence.

PROVIDER: EGAS00001004769 | EGA |

REPOSITORIES: EGA

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Single cell RNA-sequencing (scRNA-seq) of the human hematopoietic stem cell compartment (CD34+CD38-CD45RA-)

Project description:To identify a trajectory of human hematopoietic stem cell (HSC) activation we performed scRNAseq of CD34+CD38—CD45RA—cells from G-CSF-mobilized peripheral blood from 3 donors. We show that INKA1 and CDK6/PAK4 expressing single cells segregate across alternative states of quiescence.

2021-03-25 | GSE148884 | GEO

RNA Sequencing of LT-HSCs and ST-HSCs from healthy adult human Bone Marrow

Project description:The transcriptome of LT-HSC (CD34+CD38-CD45RA+CD90+CD49f+) and ST-HSC (CD34+CD38-CD45RA+CD90-CD49f-) from healthy adult human Bone Marrow Cells were assessed by RNA-seq.

2022-04-06 | E-MTAB-9892 | biostudies-arrayexpress

Single Cell analysis of the two Human Hematopoietic Stem Cell Populations

Project description:Single cell transcriptomic profiling (sc RNA-seq) of the two Human Hematopoietic Stem Cell Populations: CD34+CD38-CD45RA-CD90+CD49f+ (hereafter CD90+CD49f+) and CD34+CD38-CD45RA-CD90-CD49f+ (hereafter CD90-CD49f+)

2021-07-01 | GSE154931 | GEO

Bulk ATAC-Seq on sorted cord blood hematopoietic populations

Project description:Bulk ATAC-Seq was performed on human cord-blood derived hematopoietis popuations using the following markers: LT-HSC; CD34+CD38-CD45RA-CD90+CD49f+ ST-HSC; CD34+CD38-CD45RA-CD90-CD49f- MLP; CD34+CD38-CD45RA+ CMP; CD34+CD38+ CD10-CD45RA-Flt3+CD71- GMP; CD34+CD38+ CD10-CD45RA+ CD71- MEP; CD34+CD38+ CD10-CD45RA-Flt3-CD71+ T cell; CD3+CD19- B cell; CD19+CD3- NK cell; CD56+CD3-CD19-CD14- Granulocyte; CD14+CD15+CD3-CD19- Monocyte; CD14+CD15-CD3-CD19- DC; CD11c+CD45+ Erythroid cell; CD71+GPA+

| EGAS00001004742 | EGA

Transcriptome of cord blood derived subsets of human haematopoietic stem cells, LMPP and MLP myelo-lymphoid restricted progenitors [49f+ Subset2, Subset2, LMPPs, MLPs]

Project description:Integration of index sorting and single cell functional assays allowed identification of novel haematopoietic stem cell (HSC) and multiprogenitor subsets (MPP) that differ in their lineage differentiation potential in vitro and in vivo, cell cycle properties and long-term repopulation capacity in the NSG xenograft model. Here we report single cell transcriptomes of CD49f+ HSCs as well as those of CD49f+ Subset1 (CD19- CD38- CD45RA- CD90+ CD49f+ CD34lo CLEC9Ahi, Myelo-erythroid-skewed in vitro but Lymphoid-competent) and CD49f+ Subset2 cells (CD19- CD38- CD45RA- CD90+ CD49f+ CD34hi CLEC9Alo, Myelo-Lymphoid competent but Erythroid-deficient). We also report bulk transcriptomes of pools of 20 cells from HSC/MPP Subset1 (CD19- CD38- CD45RA- CD34lo CLEC9Ahi) and HSC/MPP Subset2 (CD19- CD38- CD45RA- CD34hi CLEC9Alo). Altogether these data show a diffuse transcriptional landscape of the CD49f+ HSC compartment, which is polarised along an axis that separates Myelo-Erythroid and Myelo-Lymphoid lineage-priming. Consistently with their differentiation capacity in vitro, CD49f+ Subset1 cells cluster at the Myelo-Erythroid end of the landscape, while CD49f+ Subset2 cells cluster at the Myelo-Lymphoid end. In addtion, these lineage-priming signatures were found to be more marked in HSC/MPP Subset1 and HSC/MPP Subset2, than in the equivalent CD49f+ subsets. In conclusion, 49f+ Subset1 and 49f+ Subset2 populations have activated distinct transcriptional lineage-priming programmes corresponding to the phenotypic lineage-skewing observed in vitro, that then become reinforced within the broader HSC/MPP pool. Altogether our data shows that lineage-priming and lineage-restriction programmes are initially established within the CD49f+ HSC subset in humans.

2018-09-14 | GSE115639 | GEO

Single cell transcriptomes of cord blood derived subsets of human haematopoietic stem cells, LMPP and MLP myelo-lymphoid restricted progenitors [49f+ HSCs, 49f+ Subset1, 49f+ Subset2, Subset1, Subset2]

2018-09-14 | GSE104995 | GEO

Isolation of single human hematopoietic stem cells capable of long-term multilineage engraftment

Project description:Gene expression profiling was used to investigate the relationship between human cord blood hematopoietic stem cells (HSC) and multipotent progenitors described in the study. Genes more highly expressed in HSC may be associated with stem cell function and self-renewal. Total RNA was obtained from flow-sorted populations based on the cell surface expression of CD34, CD38, CD45RA, Thy1, and CD49f.

2011-07-31 | E-GEOD-29105 | biostudies-arrayexpress

Profiling of CD34+ hair follicle stem cells

Project description:RNA was isolated from fluorescence activated cell sorted (FACS) CD49f+/CD34+/GFP- hair follicle stem cells after hair follicle keratinocyte isolation from the skin of age matched saline or diphtheria toxin (DT) treated Lgr5-DTReGFP mice. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0009986

2017-05-29 | GSE86962 | GEO

Mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+)

Project description:Mouse back skin was disassociated to single cells, sorted by cell surface markers and tested by microarrray To compare the gene expression of mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+) xx Bald scalp retains hair follicle stem cells but lacks CD200-rich and CD34-positive hair follicle progenitor cells Androgenetic alopecia (AGA) or common baldness results from a marked decrease in hair follicle size. This miniaturization may be related to loss of hair follicle stem or progenitor cells. To test this hypothesis, we analyzed bald and non-bald scalp from the same individuals for the presence of hair follicle stem and progenitor cells using flow cytometry to quantitate cells expressing CYTOKERATIN 15 (KRT15), CD200, CD34 and ALPHA-6-INTEGRIN (ITGA6). High levels of KRT15 expression correlated with stem cell properties of small cell size and quiescence. Cells with the highest level of KRT15 expression were maintained in bald scalp; however, distinct populations of CD200high ITGA6high cells and CD34-positive cells were markedly diminished. Consistent with a progenitor cell phenotype, the diminished populations localized closely to the stem-cell rich bulge area but were larger and more proliferative than the bulge stem cells. In functional assays, analogous CD200 high /Itga6 high cells from murine hair follicles were multipotent and generated new hair follicles in skin reconstitution assays. These findings suggest that a defect in stem cell activation plays a role in the pathogenesis of AGA. 4 independent biologic replicates (each pooled from 3 distinct mice) were sorted for Mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+)

2011-01-12 | E-GEOD-21568 | biostudies-arrayexpress

G-CSF plus AMD3100 mobilization in adult patients with beta-thalassemia results in effective mobilization of CD34+ cells with primitive signatures

Project description:Successful gene therapy for Î²-thalassemia requires optimal numbers of autologous gene-transduced hematopoietic progenitors stem cells (HPSC) with high repopulating capacity and mobilization is the optimal approach for achieving this goal. We performed a clinical study to investigate safety and efficacy of G-CSF+AMD3100 mobilization in four adult with Î²-thalassemia to reach a cell dose of â¥8x106 CD34+cells/Kg and to characterize the CD34+cell populations mobilized before and after plerixafor administration. The procedure resulted well-tolerated in all patients. Three of the four patients reached the target cell dose or more in single-apheresis collections, even one patient where a significant dose reduction of G-CSF was imposed due to early hyperleukocytosis. A significant increase in the number of circulating CD34+ cells/Âµl (12.1Â± 8.2 fold), and in the frequency of the more primitive CD34+/CD133+/CD38- cells (1.7Â±0.7 fold) was unanimously observed after AMD3100 addiction. Intraindividual comparison of gene expression profile of CD34+ cells mobilized with G-CSF versus G-CSF+ AMD3100 highlighted a different expression pattern of genes involved in the processes of HSC/stroma adhesion, homing, cell motility and transcription regulation. Overall, the safety profile, the yield, and the expression of genes that potentially promote superior engraftment depict G-CSF+AMD3100 mobilized HSC as optimal graft source for Î²-thalassemia gene therapy One experiment performed on purified CD34+ PBPC of the same patient 1 after mobilization with G-CSF + Plerixafor , and after mobilization with G-CSF alone. Universal Human Reference RNA (Stratagene, La Jolla, CA), was used as reference.

2017-04-28 | E-GEOD-70219 | biostudies-arrayexpress

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