Most primitive Human Hematopoietic Stem Cell Populations
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ABSTRACT:
PROVIDER: PRJNA646740 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR12242170_1.fastq.gz | Fastqsanger.gz | |||
| SRR12242170_2.fastq.gz | Fastqsanger.gz | |||
| SRR12242171_1.fastq.gz | Fastqsanger.gz | |||
| SRR12242171_2.fastq.gz | Fastqsanger.gz | |||
| SRR12242172_1.fastq.gz | Fastqsanger.gz |
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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:Gene expression studies from hematopoietic stem cell (HSC) populations purified to variable degrees have defined a set of stemness genes. The present study describes the construction and comparative molecular analysis of l-phage cDNA libraries from highly purified primitive HSCs (PHSCs) which retained their long term repopulating activities (LTRAs), and from maturing HSCs (MHSCs) which were largely depleted of LTRAs. Library inserts were amplified and tagged by a T7 RNA polymerase promoter and used to generate biotinylated cRNA for Microarray hybridization. Microarray analysis of the libraries confirmed previous results but also revealed an unforseen preferential expression of translation and metabolism associated genes in the PHSCs. Therefore these data indicate that HSCs are quiescent only in regard of proliferative activities, but are in a state of readiness to provide the metabolic and translational activities required following induction of proliferation by factors which induce differentiation and exit from the HSC pool. We used microarrays to detail the global programme of gene expression distinguishing primitive and maturing hematopoietic stem cells from mouse bone marrow and identified distinct classes of up- and down-regulated genes. Experiment Overall Design: To compare the transcriptosomes of primitive and maturing hematopoietic stem cells from mouse bone marrow, cDNA libraries were generated from RNA isolated from highly purified stem cell populations and used to generate biotinylated cRNA for Affymetrix microarray analysis.
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Project description:Single cell transcriptomic profiling (sc RNA-seq) of the most primitive Human Hematopoietic Stem Cell Population described: CD34+CD38-CD45RA-CD49f+EPCR+ (hereafter EPCR+).
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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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Project description:Gene expression studies from hematopoietic stem cell (HSC) populations purified to variable degrees have defined a set of stemness genes. The present study describes the construction and comparative molecular analysis of l-phage cDNA libraries from highly purified primitive HSCs (PHSCs) which retained their long term repopulating activities (LTRAs), and from maturing HSCs (MHSCs) which were largely depleted of LTRAs. Library inserts were amplified and tagged by a T7 RNA polymerase promoter and used to generate biotinylated cRNA for Microarray hybridization. Microarray analysis of the libraries confirmed previous results but also revealed an unforseen preferential expression of translation and metabolism associated genes in the PHSCs. Therefore these data indicate that HSCs are quiescent only in regard of proliferative activities, but are in a state of readiness to provide the metabolic and translational activities required following induction of proliferation by factors which induce differentiation and exit from the HSC pool. We used microarrays to detail the global programme of gene expression distinguishing primitive and maturing hematopoietic stem cells from mouse bone marrow and identified distinct classes of up- and down-regulated genes. Keywords: Adult hematopoietic stem cell populations
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