2D-DIGE Proteomic Analysis of Mesenchymal Stem Cell Cultured on the Elasticity-tunable Hydrogels

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  • Kuboki Thasaneeya
    Institute for Materials Chemistry and Engineering, Division of Biomolecular Chemistry, Kyushu University Institute for Materials Chemistry and Engineering, Division of Biomolecular Chemistry, Kyushu University
  • Kantawong Fahsai
    Centre for Cell Engineering, Division of Infection and Immunity, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow Centre for Cell Engineering, Division of Infection and Immunity, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow
  • Burchmore Richard
    Sir Henry Welcome Functional Genomics Facility, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow Sir Henry Welcome Functional Genomics Facility, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow
  • Dalby Matthew J
    Centre for Cell Engineering, Division of Infection and Immunity, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow Centre for Cell Engineering, Division of Infection and Immunity, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow
  • Kidoaki Satoru
    Institute for Materials Chemistry and Engineering, Division of Biomolecular Chemistry, Kyushu University Institute for Materials Chemistry and Engineering, Division of Biomolecular Chemistry, Kyushu University

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The present study focuses on mechanotransduction in mesenchymal stem cells (MSCs) in response to matrix elasticity. By using photocurable gelatinous gels with tunable stiffness, proteomic profiles of MSCs cultured on tissue culture plastic, soft (3 kPa) and stiff (52 kPa) matrices were deciphered using 2-dimensional differential in-gel analysis (2D-DIGE). The DIGE data, tied to immunofluorescence, indicated abundance and organization changes in the cytoskeletonal proteins as well as differential regulation of important signaling-related proteins, stress-responsing proteins and also proteins involved in collagen synthesis. The major CSK proteins including actin, tubulin and vimentin of the cells cultured on the gels were remarkably changed their expressions. Significant down-regulation of α-tubulin and β-actin can be observed on gel samples in comparison to the rigid tissue culture plates. The expression abundance of vimentin appeared to be highest in the MSCs cultured on hard gels. These results suggested that the substrate stiffness significantly affects expression balances in cytoskeletal proteins of MSCs with some implications to cellular tensegrity.

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