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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Article first published online: 22 OCT 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 11, pages 2423–2436, November 2012
How to Cite
Turco, M. Y., Furia, L., Dietze, A., Fernandez Diaz, L., Ronzoni, S., Sciullo, A., Simeone, A., Constam, D., Faretta, M. and Lanfrancone, L. (2012), Cellular Heterogeneity During Embryonic Stem Cell Differentiation to Epiblast Stem Cells Is Revealed by the ShcD/RaLP Adaptor Protein. STEM CELLS, 30: 2423–2436. doi: 10.1002/stem.1217
Author contributions: M.Y.T.: conception and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; L.F., S.R., and A.S.: collection of data; A.D.: collection and assembly of data; L.F.D.: interpretation of data; A.S. and D.C.: collection and assembly of data and data analysis and interpretation; M.F.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; L.L.: conception and design, manuscript writing, financial support, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS September 4, 2012. available online without subscription through the open access option.
- Issue published online: 22 OCT 2012
- Article first published online: 22 OCT 2012
- Accepted manuscript online: 4 SEP 2012 10:01AM EST
- Manuscript Accepted: 28 JUL 2012
- Manuscript Received: 28 NOV 2011
- Embryonic stem cells;
- Epiblast stem cells;
The Shc family of adaptor proteins are crucial mediators of a plethora of receptors such as the tyrosine kinase receptors, cytokine receptors, and integrins that drive signaling pathways governing proliferation, differentiation, and migration. Here, we report the role of the newly identified family member, ShcD/RaLP, whose expression in vitro and in vivo suggests a function in embryonic stem cell (ESC) to epiblast stem cells (EpiSCs) transition. The transition from the naïve (ESC) to the primed (EpiSC) pluripotent state is the initial important step for ESCs to commit to differentiation and the mechanisms underlying this process are still largely unknown. Using a novel approach to simultaneously assess pluripotency, apoptosis, and proliferation by multiparameter flow cytometry, we show that ESC to EpiSC transition is a process involving a tight coordination between the modulation of the Oct4 expression, cell cycle progression, and cell death. We also describe, by high-content immunofluorescence analysis and time-lapse microscopy, the emergence of cells expressing caudal-related homeobox 2 (Cdx2) transcription factor during ESC to EpiSC transition. The use of the ShcD knockout ESCs allowed the unmasking of this process as they presented deregulated Oct4 modulation and an enrichment in Oct4-negative Cdx2-positive cells with increased MAPK/extracellular-regulated kinases 1/2 activation, within the differentiating population. Collectively, our data reveal ShcD as an important modulator in the switch of key pathway(s) involved in determining EpiSC identity. STEM CELLS2012;30:2423–2436