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Synergistic effect of ascorbic acid and collagen addition on the increase in type 2 collagen accumulation in cartilage-like MSC sheet

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Abstract

Aiming to increase the content of type 2 collagen in scaffold-free cartilage-like cell sheets prepared using human bone marrow mesenchymal stem cells, the effect of several kinds of additives in a chondrogenic medium was investigated. Addition of ascorbic acid 2 phosphate (VCP) at a high concentration (250 µg/ml) and type 1 atelocollagen (5 µg/ml) increased the accumulation of type 2 collagen by fourfold and twofold, respectively. On the other hand, an antioxidant, glutathione showed no such effect. The synergistic effect of VCP and type 1 atelocollagen resulted in an eightfold increase in the accumulation level of type 2 collagen. Furthermore, the gene expression level of type 2 collagen increased and that of matrix metalloproteinase-13 (MMP-13) decreased to approximately one-third of the control. The increase in type 2 collagen accumulation in the scaffold-free cartilage-like cell sheet might be due to not only the enhancement of the synthesis but also the suppression of the degradation of type 2 collagen by MMP-13.

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Authors and Affiliations

  1. Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Kita-ku, N13W8, Sapporo, 060-8628, Japan

    Yasushi Sato & Mutsumi Takagi

  2. School of Health and Sports Sciences, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo, 663-8558, Japan

    Hisashi Mera

  3. Foundation for Biomedical Research and Innovation, International Medical Device Alliance, 1-6-5, Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan

    Hisashi Mera

  4. Department of Joint Replacement and Tissue Engineering, Graduate School of Medicine, Hokkaido University, Kita-ku, N15W7, Sapporo, 060-8638, Japan

    Tokifumi Majima

  5. Department of Orthopaedic Surgery, Graduate School of Medicine, Hokkaido University, Kita-ku, N15W7, Sapporo, 060-8638, Japan

    Daisuke Takahashi & Norimasa Iwasaki

  6. Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 739-8553, Japan

    Shigeyuki Wakitani

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  1. Yasushi Sato
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  2. Hisashi Mera
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  7. Mutsumi Takagi
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Correspondence to Mutsumi Takagi.

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Sato, Y., Mera, H., Takahashi, D. et al. Synergistic effect of ascorbic acid and collagen addition on the increase in type 2 collagen accumulation in cartilage-like MSC sheet. Cytotechnology 69, 405–416 (2017). https://doi.org/10.1007/s10616-015-9924-3

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  • DOI: https://doi.org/10.1007/s10616-015-9924-3

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