| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Embryo |
Development and Differentiation Laboratory,3 Developmental Biology Department, Insect and Animal Sciences Division, National Institute of Agrobiological Sciences, Ibaraki, 305-8602, Japan
Reproductive Cell Biology Laboratory,4 Department of Animal Breeding and Reproduction, National Institute of Livestock and Grassland Science, Ibaraki, 305-0901, Japan
We examined the expression of cell-surface markers on subpopulations of mouse embryonic stem (ES) cells to identify those that were associated with cells that had the highest pluripotency. Flow cytometry analysis revealed a 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 a high level of PECAM- 1, and reversible repopulation was observed between PECAM- 1+SSEA-1+ and PECAM-1+SSEA-1– cells. The ES cells carrying the lacZ gene were sorted into three subpopulations: PECAM- 1–SSEA-1–, PECAM-1+SSEA-1–, and PECAM-1+SSEA-1+. Quantitative reverse transcription–polymerase chain reaction 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 eight-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, many PECAM-1+SSEA- 1+ cells were found in the epiblast, but few ß-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.
2 Correspondence: Tomoyuki Tokunaga, Development and Differentiation Laboratory, Developmental Biology Department, Insect and Animal Sciences Division, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki, 305-8602, Japan. FAX: 81 298 38 7383; tom;caaffrc.go.jp
This article has been cited by other articles:
|
|
 |
|
  Y. Toyooka, D. Shimosato, K. Murakami, K. Takahashi, and H. Niwa Identification and characterization of subpopulations in undifferentiated ES cell culture Development, March 1, 2008; 135(5): 909 - 918. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Furusawa, M. Ikeda, F. Inoue, K. Ohkoshi, T. Hamano, and T. Tokunaga Gene Expression Profiling of Mouse Embryonic Stem Cell Subpopulations Biol Reprod, October 1, 2006; 75(4): 555 - 561. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Enver, S. Soneji, C. Joshi, J. Brown, F. Iborra, T. Orntoft, T. Thykjaer, E. Maltby, K. Smith, R. A. Dawud, et al. Cellular differentiation hierarchies in normal and culture-adapted human embryonic stem cells Hum. Mol. Genet., November 1, 2005; 14(21): 3129 - 3140. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Palmqvist, C. H. Glover, L. Hsu, M. Lu, B. Bossen, J. M. Piret, R. K. Humphries, and C. D. Helgason Correlation of Murine Embryonic Stem Cell Gene Expression Profiles with Functional Measures of Pluripotency Stem Cells, May 1, 2005; 23(5): 663 - 680. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
