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DOI: 10.1369/jhc.3A6241.2004
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Journal of Histochemistry and Cytochemistry
Volume 52 (11): 1447-1457, 2004
Copyright ©The Histochemical Society, Inc.

Spatial Distribution and Initial Changes of SSEA-1 and Other Cell Adhesion-related Molecules on Mouse Embryonic Stem Cells Before and During Differentiation

Li Cui, Kohei Johkura, Fengming Yue, Naoko Ogiwara, Yasumitsu Okouchi, Kazuhiko Asanuma and Katsunori Sasaki

Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine (LC,FY,KS), and Department of Anatomy and Organ Technology, Shinshu University School of Medicine (KJ,NO,YO,KA,KS), Matsumoto, Japan

Correspondence to: Li Cui, Inst. of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan. E-mail: cui{at}sch.md.shinshu-u.ac.jp

We examined the distribution of cell adhesion-related molecules (CAMs) among mouse embryonic stem (ES) cells and the spatial distribution on cell surfaces before and during differentiation. The cell–cell heterogeneity of SSEA-1, PECAM-1, and ICAM-1 among the undifferentiated cells in the ES cell colonies was evident by immunohistochemistry and immuno-SEM, supporting the flow cytometry findings. In contrast, most undifferentiated ES cells strongly expressed CD9. SSEA-1 was located preferentially on the edge of low protuberances and microvilli and formed clusters or linear arrays of 3–20 particles. PECAM-1 and ICAM-1 were randomly localized on the free cell surfaces, whereas CD9 was preferentially localized on the microvilli or protuberances, especially in the cell periphery. Both the SSEA-1+ fraction and the SSEA-1– fraction of magnetic cell sorting (MACS) formed undifferentiated colonies after plating. Flow cytometry showed that these populations reverted separately again to a culture with a mixed phenotype. Differentiation induced by retinoic acid downregulated the expression of all CAMs. Immuno-SEM showed decreases of SSEA-1 in the differentiated ES cells, although some clustering still remained. Our findings help to elucidate the significance of these molecules in ES cell maintenance and differentiation and suggest that cell surface antigens may be useful for defining the phenotype of undifferentiated and differentiated ES cells. (J Histochem Cytochem 52:1447–1457, 2004)

Key Words: embryonic stem cells • SSEA-1 • cell adhesion-related molecules • immuno-SEM and -TEM • flow cytometry • retinoic acid


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