Cytocompatibility of Lactic Acid-extracted Barramundi Skin Collagen for Cell Culture Applications

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  • MEN Yali
    Department of Histology and Embryology, Shinshu University School of Medicine
  • MA Chuang
    Shinshu University Interdisciplinary Cluster for Cutting Edge Research, Institute for Biomedical Sciences Biomedical Engineering Division, Graduate School of Medicine, Science and Technology, Shinshu University
  • TOMOTSUNE Daihachiro
    Department of Histology and Embryology, Shinshu University School of Medicine Shinshu University Interdisciplinary Cluster for Cutting Edge Research, Institute for Biomedical Sciences
  • YUE Fengming
    Department of Histology and Embryology, Shinshu University School of Medicine Shinshu University Interdisciplinary Cluster for Cutting Edge Research, Institute for Biomedical Sciences
  • JOHKURA Kohei
    Department of Histology and Embryology, Shinshu University School of Medicine

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説明

Fish collagen is becoming a potent material in many fields including the biomedical field by substituting for domestic animals that may cause zoonotic diseases. In this study, the skin of farmed barramundi (Lates calcarifer) was employed as a collagen material, and the affinity of this collagen for live cells was assessed. Four extraction solutions for the skin collagen were compared : acetic acid with/without pepsin as conventionally used, and alternatively lactic acid with/without pepsin. Among the resulting collagens, lactic acid-extracted collagen (LC) showed a meshwork of relatively fine fibers after lyophilization in scanning electron microscopy and a hydroxyproline content of 7.7％ in amino acid analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that this skin collagen was type I with α1 and α2 chains, and these chains had relatively close molecular weight as compared with those of porcine collagen. The viability of mouse fibroblasts exposed to LC at various concentrations for 24 hours was maintained at ≥ 93％, demonstrating its low cytotoxicity. In a wound healing assay using each collagen in the culture medium, LC supported more active motility of mouse fibroblasts than collagens obtained by the other extraction methods. Collectively, these results suggest that barramundi skin collagen extracted by lactic acid has suitable properties in cellular activities, exhibiting advantages over the conventional methods, and is expected to be a useful material for cell culture applications.

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