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Heat shock pretreatment enhances porcine myoblasts survival after autotransplantation in intact skeletal muscle

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Abstract

Myoblast transplantation (MT) is a cell-based gene therapy treatment, representing a potential treatment for Duchenne muscular dystrophy (DMD), cardiac failure and muscle trauma. The rapid and massive death of transplanted cells after MT is considered as a major hurdle which limits the efficacy of MT treatment. Heat shock proteins (HSPs) are overexpressed when cells undergo various insults. HSPs have been described to protect cells in vivo and in vitro against diverse insults. The aim of our study is to investigate whether HSP overexpression could increase myoblast survival after autotransplantation in pig intact skeletal muscle. HSP expression was induced by warming the cells at 42°C for 1 h. HSP70 expression was quantified by Western blot and flow cytometry 24 h after the treatment. To investigate the myogenic characteristics of myoblasts, desmin and CD56 were analysed by Western blot and flow cytometry; and the fusion index was measured. We also quantified cell survival after autologous transplantation in pig intact skeletal muscle and followed cell integration. Results showed that heat shock treatment of myoblasts induced a significative overexpression of the HSP70 (P < 0.01) without loss of their myogenic characteristics as assessed by FACS and fusion index. In vivo (n=7), the myoblast survival rate was not significantly different at 24 h between heat shock treated and nontreated cells (67.69% ± 8.35% versus 58.79% ± 8.35%, P > 0.05). However, the myoblast survival rate in the heat shocked cells increased by twofold at 48 h (53.32% ± 8.22% versus 28.27% ± 6.32%, P < 0.01) and more than threefold at 120 h (26.33% ± 5.54% versus 8.79% ± 2.51%, P < 0.01). Histological analysis showed the presence of non-heat shocked and heat shocked donor myoblasts fused with host myoblasts. These results suggested that heat shock pretreatment increased the HSP70 expression in porcine myoblasts, and improved the survival rate after autologous transplantation. Therefore, heat shock pretreatment of myoblast in vitro is a simple and effective way to enhance cell survival after transplantation in pig. It might represent a potential method to overcome the limitations of MT treatment.

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Affiliated Zhongshan Hospital of Dalian University, Dalian, 116001, China

    Yang Sheng

  2. Department of Orthopedic Surgery, Geneva Faculty of Medicine and University Hospital, CH-1211, Geneva, Switzerland

    Yang Sheng, Thomas Laumonier & Jacques Menetrey

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  1. Yang Sheng
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  2. Thomas Laumonier
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  3. Jacques Menetrey
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Supported by the Swiss National Science Foundation (NRP 46 no. 4046-058639)

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Yang, S., Laumonier, T. & Menetrey, J. Heat shock pretreatment enhances porcine myoblasts survival after autotransplantation in intact skeletal muscle. SCI CHINA SER C 50, 438–446 (2007). https://doi.org/10.1007/s11427-007-0065-6

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  • DOI: https://doi.org/10.1007/s11427-007-0065-6

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