西亚试剂：Heparan Sulfate Regulates Self-renewal and Pluripotency of

Heparan Sulfate Regulates Self-renewal and Pluripotency of Embryonic Stem Cells*

Norihiko Sasaki ${ddagger}$ $§$ , Kazuhiko Okishio ${ddagger}$ , Kumiko Ui-Tei¶, Kaoru Saigo¶, Akiko Kinoshita-Toyoda $§$ ||, Hidenao Toyoda $§$ ||, Tomoaki Nishimura** ${ddagger}$ ${ddagger}$ , Yasuo Suda $§$ ** ${ddagger}$ ${ddagger}$ , Michiko Hayasaka $§$ $§$ , Kazunari Hanaoka $§$ $§$ , Seiji Hitoshi¶¶, Kazuhiro Ikenaka¶¶, and Shoko Nishihara ${ddagger}$ $§$ 1

From the ${ddagger}$ Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, the ¶Department of Biophysics and Biochemistry, Graduate School of Science, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, the ||Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi, Inage, Chiba 263-8522, the **Department of Nanostructure and Advanced Materials, Graduate School of Science and Engineering, Kagoshima University, 1-21-40 Kohrimoto, Kagoshima 890-0065, ${ddagger}$ ${ddagger}$ Sudx-Biotec Corporation, KIBC 461, 5-5-2, Minatojima-minami, Chuo-ku, Kobe 650-0047, $§$ $§$ Laboratory of Molecular Embryology, Department of Bioscience, Kitasato University School of Science, 1-15-1 Kitasato, Sagamihara, Kanagawa 228, ¶¶Division of Neurobiology and Bioinformatics, National Institute for Physiological Sciences, 38 Nishigonaka Myodaiji, Okazaki, Aichi 444-8585, and $§$ Core Research for Evolutional Science and Technology of Japan Science and Technology Agency, Kawaguchi Center Building, 4-1-8, Hon-cho, Kawaguchi, Saitama 332-0012, Japan

Embryonic stem (ES) cell self-renewal and pluripotency are maintained by several signaling cascades and by expression of intrinsic factors, such as Oct3/4 and Nanog. The signaling cascades are activated by extrinsic factors, such as leukemia inhibitory factor, bone morphogenic protein, and Wnt. However, the mechanism that regulates extrinsic signaling in ES cells is unknown. Heparan sulfate (HS) chains are ubiquitously present as the cell surface proteoglycans and are known to play crucial roles in regulating several signaling pathways. Here we investigated whether HS chains on ES cells are involved in regulating signaling pathways that are important for the maintenance of ES cells. RNA interference-mediated knockdown of HS chain elongation inhibited mouse ES cell self-renewal and induced spontaneous differentiation of the cells into extraembryonic endoderm. Furthermore, autocrine/paracrine Wnt/β-catenin signaling through HS chains was found to be required for the regulation of Nanog expression. We propose that HS chains are important for the extrinsic signaling required for mouse ES cell self-renewal and pluripotency.

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