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Newly defined human hematopoietic stem cell populations

ABSTRACT: Transcriptomic profiling (RNA-seq) of primitive human progenitor populations CD34+CD38-CD45RA-CD90-EPCR-; hereafter CD90-EPCR- MPP.

ORGANISM(S): Homo sapiens

PROVIDER: GSE197079 | GEO | 2022/02/23

REPOSITORIES: GEO

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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+)

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Project description:Purpose: to compare the transcriptomic profile of the two most primitive human EPCR+ stem and CD90+EPCR- progenitor populations. Methods: cells were obtained from human cord-blood and flow-sorted using the surface antigens described. 5 EPCR+ and 4 CD90+EPCR- biological replicates were used to extract total RNA and quality was verified (RIN >8). Libraries were prepared using with 20-25 ng/sample of starting RNA. All samples were sequenced two independent times. Conclusions: this study represents the first analysis of transcriptomes from the two most primitive human stem/progenitor populations, with biologic replicates, 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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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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Most primitive Human Hematopoietic Stem Cell Populations

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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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]

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