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Growth arrest of vascular smooth muscle cells in suspension culture using low-acyl gellan gum

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Abstract

The proliferation of vascular smooth muscle cells (SMCs) causes restenosis in biomaterial vascular grafts. The purposes of this study were to establish a suspension culture system for SMCs by using a novel substrate, low-acyl gellan gum (GG) and to maintain SMCs in a state of growth inhibition. When SMCs were cultured in suspension with GG, their proliferation was inhibited. Their viability was 70% at day 2, which was maintained at more than 50% until day 5. In contrast, the viability of cells cultured in suspension without GG was 5.6% at day 2. By cell cycle analysis, the ratio of SMCs in the S phase when cultured in suspension with GG was lower than when cultured on plastic plates. In SMCs cultured in suspension with GG, the ratio of phosphorylated retinoblastoma (Rb) protein to Rb protein was decreased and p27Kip1 expression was unchanged in comparison with SMCs cultured on plastic plates. In addition, SMCs could be induced to proliferate again by changing the culture condition from suspension with GG to plastic plates. These results suggest that our established culturing method for SMCs is useful to maintain SMCs in a state of growth inhibition with high viability.

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Acknowledgments

This study was performed under a partnership of collaboration between Chiba University and Nissan Chemical Industries Ltd. The investigations using animals described in this report conformed to the guidelines of the Animal Investigation Committee of Chiba University (Chiba, Japan).

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Authors and Affiliations

  1. Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan

    Tomomi Natori, Masachika Fujiyoshi & Itsuko Ishii

  2. Division of Pharmacy, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8677, Japan

    Masashi Uchida & Itsuko Ishii

  3. Biological Research Laboratories, Nissan Chemical Industries Ltd., 1470 Shiraoka, Shiraoka, Saitama, 369-0305, Japan

    Natsuki Abe & Tatsuro Kanaki

  4. Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan

    Yasunori Fukumoto

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  1. Tomomi Natori
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  2. Masachika Fujiyoshi
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  3. Masashi Uchida
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  4. Natsuki Abe
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  6. Yasunori Fukumoto
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  7. Itsuko Ishii
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Correspondence to Itsuko Ishii.

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Editor: Tetsuji Okamoto

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Natori, T., Fujiyoshi, M., Uchida, M. et al. Growth arrest of vascular smooth muscle cells in suspension culture using low-acyl gellan gum. In Vitro Cell.Dev.Biol.-Animal 53, 191–198 (2017). https://doi.org/10.1007/s11626-016-0098-x

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  • DOI: https://doi.org/10.1007/s11626-016-0098-x

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