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First published online May 11, 2006
Stem Cells Vol. 24 No. 8 August 2006, pp. 1937 -1945
doi:10.1634/stemcells.2005-0067; www.StemCells.com
© 2006 AlphaMed Press

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EMBRYONIC STEM CELLS: CHARACTERIZATION SERIES

Quantitative Oct4 Overproduction in Mouse Embryonic Stem Cells Results in Prolonged Mesoderm Commitment During Hematopoietic Differentiation In Vitro

Valérie Camara-Clayette, Françoise Le Pesteur, William Vainchenker, Françoise Sainteny

INSERM U790, IFR54, Institut Gustave Roussy, Villejuif cedex, France

Key Words. Embryonic stem cell • Hematopoiesis • Cell differentiation • Transcription factor • Mesoderm • Gene expression

Correspondence: Valérie Camara-Clayette, Ph.D., Institut Gustave Roussy, INSERM U790, PR1, 94805 Villejuif cedex, France. Telephone: +33 1 42 11 42 33; Fax: +33 1 42 11 54 90; e-mail: vclayet{at}igr.fr

Received February 17, 2005; accepted for publication May 4, 2006.
First published online in STEM CELLS EXPRESS   May 11, 2006.



The Oct4 transcription factor is essential for the self-renewal and pluripotency of embryonic stem cells (ESCs). Oct4 level also controls the fate of ESCs. We analyzed the effects of Oct4 overproduction on the hematopoietic differentiation of ESCs. Oct4 was introduced into ESCs via a bicistronic retroviral vector, and cells were selected on the basis of Oct4 production, with Oct4+ and Oct42+ displaying twofold and three- to fourfold overproduction, respectively. Oct4 overproduction inhibited hematopoietic differentiation in a dose-dependent manner, after the induction of such differentiation by the formation of day 6 embryoid bodies (EB6). This effect resulted from defective EB6 formation rather than from defective hematopoietic differentiation. In contrast, when hematopoiesis was induced by the formation of blast colonies, the effects of Oct4 depended on the level of overproduction: twofold overproduction increased hematopoietic differentiation, whereas higher levels of overproduction markedly inhibited hematopoietic development. This increase or maintenance of Oct4 levels appears to alter the kinetics and pattern of mesoderm commitment, thereby modifying hemangioblast generation. These results demonstrate that Oct4 acts as a master regulator of ESC differentiation.




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