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A more recent version of this article appeared on April 1, 2002
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Submitted on October 31, 2001
Revised on December 14, 2001
Accepted on January 9, 2002
1 Department of Pathology, Program in Stem Cell Biology, Shands Cancer Center, University of Florida College of Medicine, Gainesville, FL 32610
2 Department of Pharmacology, University of Florida College of Medicine, Gainesville, FL 32610
3 Department of Stem Cell Regulation, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
* Corresponding author. E-mail address: terada{at}pathology.ufl.edu.
Mouse embryonic stem (ES) cells can proliferate indefinitely in an undifferentiated state in the presence of leukemia inhibitory factor (LIF), or differentiate into all three germ layers upon removal of this factor. In order to determine cellular factors associated with self-renewal of undifferentiated ES cells, we used PCR-assisted cDNA subtraction to screen genes that are expressed in undifferentiated ES cells and down-regulated after incubating these cells in a differentiation medium without LIF for 48 hrs. The mRNA expression of a tetraspanin transmembrane protein CD9 was high in undifferentiated ES cells and decreased shortly after cell differentiation. An immunohistochemical analysis confirmed that plasma membrane-associated CD9 was expressed in undifferentiated ES cells but low in the differentiated cells. Addition of LIF to differentiating ES cells re-induced mRNA expression of CD9, and CD9 expression was accompanied with a reappearance of undifferentiated ES cells. Further, activation of STAT3 induced the expression of CD9, indicating the LIF/STAT3 pathway is critical for maintaining CD9 expression. Finally, addition of anti-CD9 antibody blocked ES cell colony formation and reduced cell viability. These results indicate that CD9 may play a role in LIF-mediated maintenance of undifferentiated ES cells.
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