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BOR - Papers in Press, published online ahead of print January 21, 2004.
Biol Reprod 2004, 10.1095/biolreprod.103.024190
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Submitted October 14, 2003
Returned for revision October 29, 2003
Accepted January 12, 2004

Embryo


Embryonic Stem Cells Expressing Both Platelet Endothelial Cell Adhesion Molecule-1 and Stage-Specific Embryonic Antigen-1 Differentiate Predominantly into Epiblast Cells in a Chimeric Embryo

Tadashi Furusawa , Katsuhiro Ohkoshi , Chris Honda , Seiya Takahashi , and Tomoyuki Tokunaga *

* To whom correspondence should be addressed. E-mail: tom{at}affrc.go.jp.

Abstract
We examined the expression of cell-surface markers on subpopulations of mouse embryonic stem (ES) cells in order to identify those that were associated with cells that had the highest pluripotency. Flow cytometry analysis revealed wide variation in the expression of platelet endothelial cell adhesion molecule 1 (PECAM-1) and stage-specific embryonic antigen (SSEA)-1 in ES cells. Almost all SSEA-1+ cells expressed high level of PECAM-1, and reversible repopulation was observed between PECAM-1+SSEA-1+ and PECAM-1+SSEA-1- cells. ES cells carrying the lacZ gene were sorted into three sub populations: PECAM-1-SSEA-1-, PECAM-1+ SSEA-1- and PECAM-1+ SSEA-1+. Quantitative RT-PCR revealed a low level of Oct3/4 mRNA expression and an elevation in differentiation maker gene expression in PECAM-1- cells. To compare the pluripotency of these three subpopulations, a single cell from each was injected into 8-cell embryo and ES cells identified at later stages by X-gal staining. At the blastocyst stage, PECAM-1+ SSEA-1+/- cells were found to have differentiated into epiblast cells in high numbers. In contrast, PECAM-1- cell derivatives localized in the primitive endoderm or trophectoderm. At 6.0-7.0 days post coitum (dpc), many PECAM-1+SSEA-1+ cells were found in the epiblast, but few {beta}-gal+ cells were detected in any regions of embryos that were injected with cells from the other two populations. These results showed that the expression levels of PECAM-1 and SSEA-1 in ES cells correlated closely with their pluripotency and/or their ability to incorporate into the epiblast of chimeric embryos.

Key words: Embryo • Developmental biology • Early development


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