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Mitochondrial activity linked to CD34+CD90+EPCR+ phenotype determines the functional fitness of ex vivo expanded HSCs

ABSTRACT: Mitochondrial activity linked to CD34+CD90+EPCR+ phenotype determines the functional fitness of ex vivo expanded HSCs

PROVIDER: PRJNA648928 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12331566.fastq.gz	Fastqsanger.gz
	SRR12331567.fastq.gz	Fastqsanger.gz
	SRR12331568.fastq.gz	Fastqsanger.gz
	SRR12331569.fastq.gz	Fastqsanger.gz
	SRR12331570.fastq.gz	Fastqsanger.gz

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Mitochondrial activity linked to CD34+CD90+EPCR+ phenotype determines the functional fitness of ex vivo expanded HSCs

Project description:We report the transcritpome of purified CD34+ cells from cultures initiated with cord blood CD34+ hematopoietic stem cells that were expanded ex vivo in Stemline II media supplemented with a cytokine cocktail in the presence or absence of valproic acid for 6 days .

2020-11-20 | GSE155180 | GEO

Single cell transcriptome sequencing of expanded human hematopoieic stem/progenitor cells cultured in PCL-PVAc-PEG based 3a medium and StemSpan with UM171/SR-1 medium.

Project description:We demonstrated that chemically difined PCL-PVAc-PEG based 3a medium was superior to conventional StemSpan besed medium in expansion of human hematopoietic stem cells. In fact, more phenotypic HSCs (CD34+EPCR+CD90+CD45RA-ITGA3+) were expanded in our culture condition and higher donor chmerism was achieved in transplantation assay using humanized mice. But detail transcriptome profiles of expanded HSCs was unknown and therefore single-cell RNA sequencing was done to reveal it. As a result, expanded cells in our medium had more HSPC featured cells. Interestingly, erythroid/megakaryocyte progenitors was more expanded in our condition. On the other hands, other culture condition made more myeloid progenitor cells but few HSPC featured cells. This result support the idea that our new medium are able to selectively expand HSCs.

2022-11-13 | GSE192519 | GEO

Limited Mitochondrial Activity Coupled With Strong Expression of CD34, CD90 and EPCR Determines the Functional Fitness of <i>ex vivo</i> Expanded Human Hematopoietic Stem Cells.

Project description:Ex vivo expansion strategies of human hematopoietic stem cell (HSC) grafts with suboptimal stem cell dose have emerged as promising strategies for improving outcomes of HSC transplantation in patients with hematological malignancies. While exposure of HSCs to ex vivo cultures expands the number of phenotypically identifiable HSCs, it frequently alters the transcriptomic and metabolic profiles, therefore, compromising their long-term (LT) hematopoietic reconstitution capacity. Within the heterogeneous pool of expanded HSCs, the precise phenotypic, transcriptomic and metabolic profile and thus, the identity of HSCs that confer LT repopulation potential remains poorly described. Utilizing valproic acid (VPA) in ex vivo cultures of umbilical cord blood (UCB)-CD34+ cells, we demonstrate that expanded HSCs phenotypically marked by expression of the stem cell markers CD34, CD90 and EPCR (CD201) are highly enriched for LT-HSCs. Furthermore, we report that low mitochondrial membrane potential, and, hence, mitochondrial activity distinguishes LT-HSCs within the expanded pool of phenotypically defined HSCs. Remarkably, such reduced mitochondrial activity is restricted to cells with the highest expression levels of CD34, CD90 and EPCR phenotypic markers. Together, our findings reveal that high expression of CD34, CD90 and EPCR in conjunction with low mitochondrial activity is critical for identification of functional LT-HSCs generated within ex vivo expansion cultures.

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

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