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Hematopoietic reconstitution by multipotent adult progenitor cells: precursors to long-term hematopoietic stem cells

ABSTRACT: We used comparative transcriptome analysis to determine the relationship between rodent Multipotent Adult Progenitor Cells (MAPCs) and pluripotent embryonic stem cells (ESCs), or lineage restricted mesenchymal stem cells (MSCs) and neurospheres (NS). This comparison revealed a unique gene expression profile of MAPCs, that express transcripts of early endodermal and mesodermal but fewer ectodermal genes. In addition, MAPCs, but not MSCs or NS, express a number of genes expressed in ESCs including Oct-4 (Pou5f1) and other genes previously identified as ES cell associated transcripts and/or genes involved in maintenance of ESC selfrenewal capacity and pluripotency. However MAPCs do not express some key genes involved in maintaining selfrenewal and pluripotency in ESCs, including Nanog and Sox-2. Because MSC-like cells isolated under MAPC conditions are virtually identical to MSCs isolated in the presence of high serum, the MAPC signature is cell-type specific and not merely the result of differing culture conditions. Finally, this unique molecular signature was seen irrespective of the microarray platform used, and was highly similar for both mouse and rat MAPCs. Keywords: cell type comparison design

ORGANISM(S): Mus musculus

PROVIDER: GSE5947 | GEO | 2007/01/26

SECONDARY ACCESSION(S): PRJNA97401

REPOSITORIES: GEO

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Hematopoietic reconstitution by multipotent adult progenitor cells: precursors to long-term hematopoietic stem cells

Project description:We used comparative transcriptome analysis to determine the relationship between rodent Multipotent Adult Progenitor Cells (MAPCs) and pluripotent embryonic stem cells (ESCs), or lineage restricted mesenchymal stem cells (MSCs) and neurospheres (NS). This comparison revealed a unique gene expression profile of MAPCs, that express transcripts of early endodermal and mesodermal but fewer ectodermal genes. In addition, MAPCs, but not MSCs or NS, express a number of genes expressed in ESCs including Oct-4 (Pou5f1) and other genes previously identified as ES cell associated transcripts and/or genes involved in maintenance of ESC selfrenewal capacity and pluripotency. However MAPCs do not express some key genes involved in maintaining selfrenewal and pluripotency in ESCs, including Nanog and Sox-2. Because MSC-like cells isolated under MAPC conditions are virtually identical to MSCs isolated in the presence of high serum, the MAPC signature is cell-type specific and not merely the result of differing culture conditions. Finally, this unique molecular signature was seen irrespective of the microarray platform used, and was highly similar for both mouse and rat MAPCs. Keywords: cell type comparison design Two biological replicates were used for hybridizations. cRNA from MAPCs, ESCs, and NS were added to a universal reference cRNA.

2010-06-11 | E-GEOD-5947 | biostudies-arrayexpress

The forced expression of Nanog or Esrrb allows preserve ESC status in the absence of nucleostemin expression

2015-01-30 | GSE56797 | GEO

The forced expression of Nanog or Esrrb allows preserve ESC status in the absence of nucleostemin expression

Project description:Nucleostemin (NS) gene is known to be expressed in stem cells in general including embryonic stem cells (ESCs). Previous knockdown and knockout studies have demonstrated that NS is important for the preservation of their self-renewality and high levels of pluripotent marker gene expression in mouse ESCs. In this study, we demonstrate that the forced expression of Nanog or Esrrb, but not other pluripotency factors, made NS expression dispensable in mouse ESCs. DNA microarray data deposited here underscored the notion that both Nanog and Esrrb could rather faithfully counteract the alteration of gene expression profile caused by NS expression ablation in ESCs. NS tet off ESCs (Stem Cells 27, 1066-1076, 2009) in which NS gene was homozygously disrupted, but NS cDNA was introduced in ROSA26 gene locus together with tetracycline-off system were used as parental ESCs to examine the effect of NS expression ablation in ESCs. In this study, how alteration of gene expression profile change associated with NS expression ablation was modulated by the forced expression of either constitutively active Stat3 (Stat3mut), Nanog or Esrrb. Effect obtained by the exposure to 2i condition (medium containing 2 kinase inhibitors against MEK and GSK3b) was also examined

2015-01-30 | E-GEOD-56797 | biostudies-arrayexpress

Pramel7-Cul2 axis establishes ground-state gene expression in embryonic stem cells

Project description:Pluripotency is established in E4.5 preimplantation epiblast, the founder populations of the adult embryo. The establishment of in vitro models that closely resemble ground-state pluripotency represents an important opportunity to study early embryo development. Pramel7, a protein highly expressed in the inner cell mass (ICM) but expressed at low levels in embryonic stem cells (ESCs), was implicated in the establishment of ground-state pluripotency. Increasing Pramel7 expression in developmentally advanced ESC+serum causes global DNA hypomethylation and induces a gene expression signature close to developmental ground-state. However, how Pramel7 affects gene expression remains elusive. Here we show that Pramel7 associates and recruits to chromatin Cullin2 (Cul2), a component of Cullin2-RING E3 ubiquitin ligase complex that is implicated in proteasomal degradation of target substrates. Pramel7-Cul2 interaction is required for the establishment of ground-state gene expression signature. We show that Pramel7/Cul2 directly target many components of repressive chromatin, including components of the Nucleosome Remodelling and Deacetylase (NuRD) complex, for degradation. We identified a set of Pramel7-regulated genes that depend on Cul2 and associate with the NuRD component Chd4. The majority of these genes are upregulated in ESCs expressing Pramel7 and linked to pluripotency pathways. Finally, we show that Chd4 binding to these genes is impaired by Pramel7 in a Cul2-dependent manner. Our data link proteasome pathway to the establishment of ground-state gene expression, offering insights that could facilitate the establishment of in vitro models to reproduce the in vivo ground-state pluripotency.

2024-01-08 | PXD032184 | Pride

A balanced Oct4 protein interaction network is crucial for maintaining pluripotency

Project description:Oct4 is considered a master transcription factor for pluripotent cell self-renewal and embryo development. It primarily collaborates with other transcriptional factors or coregulators to maintain pluripotency. However, it is still unclear how Oct4 interacts with its partners. Here, we show that the Oct4 linker interface mediates competing and balanced Oct4 protein interactions which are crucial for maintaining pluripotency. Linker mutant ESCs maintain the key pluripotency genes expression, but show decreased expression of self-renewal genes and increased expression of differentiation genes which result in impaired ESCs self-renewal and early embryonic lethality. Linker mutation dose not affect Oct4 genomic binding and transactivation potential, but breaks the balanced Oct4 interactome. In mutant ESCs, the interaction between Oct4 and Klf5 was decreased, but interactions between Oct4 and Cbx1, Ctr9, Cdc73 were increased which disrupt the epigenetic state of ESCs. Overexpression of Klf5 or knockdown Cbx1, Cdc73 rescue the cellular phenotype of linker mutant ESCs by rebalancing Oct4 interactome indicating that different partners interact with Oct4 competitively. Thus, by showing how Oct4 interacts with different partners, we provide novel molecular insights to explain how Oct4 contributes to the maintenance of pluripotency.

2021-12-10 | PXD017456 | Pride

Unique Molecular Signature of Multipotent Adult Progenitor Cells (Affy)

2007-10-11 | GSE6933 | GEO

Derivation of porcine extra-embryonic endoderm cell lines reveals distinct signaling pathway and multipotency states

Project description:We detected the gene expression differences between porcine extra-embryonic endoderm cells and naïve like and primed porcine ESCs. Gene expression of porcine extra-embryonic endoderm cells was distinct with porcine naïve like and primed embryonic stem cells. Pearson correlation analysis confirmed that porcine naïve like and primed ESCs have stronger correlation than the correlation between either pXEN cells and porcine naïve like ESCs or primed ESCs. Porcine extra-embryonic endoderm cells have a distinct identity, clustering neither with porcine naïve like ESCs nor with primed ESCs. We confirmed that porcine extra-embryonic endoderm cells lack the expression of key pluripotency genes, such as Sox2, Pou5f1 and Klf4, but they expressed other pluripotency genes, such as Sall4, Lin28a and Klf5. Furthermore, porcine extra-embryonic endoderm cells robustly express Gata4, Gata6 and Sox17 as well as genes encoding ExEn-associated cell surface proteins or basement membrane components Pdgfra, Col4a2, Lama1, Lamb1, Sparc, Ihh, Hnf4a and Dab2, which is in contrast to porcine naïve like ESCs that express these genes at a low level or lack expression altogether

2021-12-01 | GSE183270 | GEO

Functional dissection of hematopoietic stem cell population by a stemness-monitoring system with NS-GFP transgene

Project description:Hematopoietic stem cells (HSCs) in a steady state are efficiently purified by the combination of several cell surface markers, however, such markers do not consistently reflect HSC activity. In this study, we successfully isolated functional HSCs by a unique stemness monitoring system using a transgenic mouse, in which green florescence protein (GFP) is driven by promoter/enhancer region of nucleostemin (NS) gene. We found that phenotypically defined long-term (LT)-HSC population exhibited the highest level of the NS-GFP intensity, whereas it was remarkably down-regulated during differentiation in vitro and in vivo. Importantly, among the LT-HSC population, the NS-GFPhigh cells significantly exhibited higher repopulating capacity than NS-GFPlow cells. Gene expression analysis revealed that 9 genes, including Vwf and Cdkn1c (p57), are highly expressed in NS-GFPhigh cells, which may represent signature of functional HSCs, i.e., stemness signature. When LT-HSCs suffered from remarkable stresses, such as transplantation or irradiation, NS-GFP intensity was downregulated, suggesting that NS-GFP accurately reflects HSC function. Finally, we found that functional HSCs were identified as NS-GFPhigh cells in CD34+ LSK cells, which has been recognized as progenitor cells without long-term reconstitution ability. Thus, high NS-GFP intensity represents stem cell characteristics in hematopoiesis and this system is useful for identification of uncharacterized HSCs.

2017-12-20 | GSE98500 | GEO

Single Cell based genomewide gene expression analysis of murine bone-marrow derived Very Small Embryonic-Like Stem Cells (VSELs)

Project description:Recently, we identified a population of Oct4+Sca-1+Lin-CD45- very small embryonic-like stem-cells (VSELs) in adult tissues. Open chromatin structure of pluripotency genes and genomic imprinting-related epigenetic mechanisms maintain pluripotency and quiescence of VSELs, respectively. However, global transcriptome signature of this rare stem-cell population remains elusive. Here, we demonstrate by genomewide gene-expression analysis with a small number of highly purified murine bone-marrow (BM)-derived VSELs, that Oct4+ VSELs i) express a similar, yet nonidentical, transcriptome as embryonic stem-cells (ESCs), ii) up-regulate cell-cycle checkpoint genes, iii) down-regulate genes involved in protein turnover and mitogenic pathways, and iv) highly express Ezh2, a polycomb group protein. Furthermore, as a result of Ezh2 overexpression, VSELs, like ESCs, exhibit bivalently modified nucleosomes (trimethylated H3K27 and H3K4) at promoters of important homeodomain-containing developmental transcription factors, thus preventing their premature transcription. Notably, spontaneous or RNA interference-enforced down-regulation of Ezh2 during VSEL differentiation removes the bivalent-domain (BD) structure, which leads to de-repression of several BD-regulated genes. Therefore, we suggest that VSELs, like other pluripotent stem-cells, maintain their pluripotent state through an Ezh2-dependent BD-mediated epigenetic mechanism.

2011-12-21 | GSE29281 | GEO

Transcription profiling of mouse multipotent adult progenitor cells

Project description:We compare the transcriptome of embryonic stem cells (ESCs), adult stem cells with apparent greater differentiation potential such as multipotent adult progenitor cells (MAPCs), mesenchymal stem cells (MSCs) and neurospheres (NS). Mouse and rat MAPCs were used in this study and two different array platforms (Affymetrix and NIA) were used for mouse samples. Experiment Overall Design: Three mRNA samples (biological replicates) per cell type taken at different passage number were compared. Cell types include mouse ESCs, mouse MSCs and three clones isolated using mouse MAPC culture condition: mMAPC-1, mMAPC-2 and mClone-3. mMAPC-1 and mClone-3 were obtained from same bone marrow isolation, mMAPC-2 was obtained in a different bone marrow isolation. Two clones derived from same rat bone marrow using rat MAPC culture conditions were compared. Three mRNA samples (biological replicates) per clone were taken at different passage numbers

2008-06-15 | E-GEOD-6933 | biostudies-arrayexpress

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