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Single Cell analysis of the two Human Hematopoietic Stem Cell Populations

ABSTRACT: 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+)

ORGANISM(S): Homo sapiens

PROVIDER: GSE154931 | GEO | 2021/07/01

REPOSITORIES: GEO

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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.

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Project description:Transcriptomic profiling (RNA-seq) of the four most primitive human stem and progenitor populations. Purpose: to compare the transcriptomic profile of the four most primitive human stem and progenitor populations (all are CD34+CD38-CD45RA-): CD90+CD49f+, CD90-CD49f+, CD90+CD49f-, CD90-CD49f-. Conclusions: previouls study of gene expression analysis of these populations were performed using microarray hence, this study represents the first analysis of transcriptomes from these populations generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles of other human blood stem/progenitor populations.

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Project description:10x single cell sequencing of LT-HSCs (CD34+CD38-CD90+CD45RA-CD49f+) from human bone marrow.

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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

Functional and transcriptional heterogeneity of human hemopoietic lympho-myeloid progenitors

Project description:Cord blood (CB) samples from normal donors were obtained with informed consent. Fresh CB samples were processed within 18-34h after collection. Mononuclear cells were isolated and CD34+ fraction was separated. CB CD34+ enriched fraction was lineage depleted by staining with purified anti-human CD2, CD3, CD4, CD7, CD8a, CD11b, CD14, CD19, CD20, CD56, CD235a followed by Qdot 605 conjugated goat F(ab')2 anti-mouse IgG (H+L). Cells were also stained with anti-human CD38-FITC, CD45RA-PE or -BV650, CD123-PE Cy7, CD90-biotin, CD34- PerCP and CD10-APC. Finally, cells were incubated with streptavidin-conjugated APC-eF780 and Hoechst 33258 (Invitrogen, final concentration: 1 g/ml). Populations were defined, as follows: HSC - Lin-CD34+CD38-CD90+CD45RA-CD10-, MPP - Lin-CD34+CD38-CD90-CD45RA-CD10-, LMPP - Lin-CD34+CD38-CD90-/loCD45RA+CD10-, MLP - Lin-CD34+CD38-CD90-/loCD45RA+CD10+, GMP - Lin-CD34+CD38+CD123+CD45RA+CD10-, CMP - Lin-CD34+CD38+CD123+CD45RA-CD10-, MEP - Lin-CD34+CD38+CD123-CD45RA-CD10-.

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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]

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 | GSE104995 | GEO

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

2018-09-14 | GSE115639 | GEO

Gene expression profiling of novel hematopoietic stem cell subpopulations defined by CD35 expression and progenitor populations in human adult hematopoiesis

Project description:Cannonical hematopoietic stem cell (HSC) population is phenotypically defined as Lin- CD34+ CD38- CD45RA- CD90+ in human adult bone marrow. We recently identified novel HSC subpopulation defined as CD35+ HSCs. We performed microarray analysi to clarify the characteristics of CD35+ HSCs in comparison with other stem/progenitor populations. As compared to CD35- HSCs and progenitor populations, CD35+ HSCs showed lower expression of cell-cycle- or lineage-affiliated genes.suggesting their dormant or immature state.

2018-04-13 | GSE113033 | GEO

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