Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

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

ABSTRACT: 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. T7-primed cDNA libraries was synthesized using FACS sorted twenty cells. T7-primed libraries were biotin-labeled using the GeneChip 3’ in vitro transcription (IVT) kit (Affymetrix), starting from ‘‘In vitro Transcription to Synthesize Labeled aRNA. Nine Samples (3 VSELs, 3 HSC, and 3 ESC-D3) are analyzed using GeneChip 3’ Mouse Genome 430 2.0 array (Affymetrix), according to the manufacturer’s instructions.

ORGANISM(S): Mus musculus

SUBMITTER: DONG-MYUNG SHIN

PROVIDER: E-GEOD-29281 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Global gene expression analysis of very small embryonic-like stem cells reveals that the Ezh2-dependent bivalent domain mechanism contributes to their pluripotent state.

Shin Dong-Myung DM Liu Rui R Wu Wan W Waigel Sabine J SJ Zacharias Wolfgang W Ratajczak Mariusz Z MZ Kucia Magda M

Stem cells and development 20120104 10

Recently, we identified a population of Oct4(+)Sca-1(+)Lin(-)CD45(-) very small embryonic-like stem cells (VSELs) in murine and human adult tissues. VSELs can differentiate in vitro into cells from all 3 germ layers and in vivo tissue-committed stem cells. Open chromatin structure of core pluripotency transcription factors (TFs) supports the pluripotent state of VSELs. However, it has been difficult to determine how primitive VSELs maintain pluripotency, owing to their limited number in adult ti ...[more]

PMID: 22023227

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Dataset Information

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

Publications

Global gene expression analysis of very small embryonic-like stem cells reveals that the Ezh2-dependent bivalent domain mechanism contributes to their pluripotent state.

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