Development of 3D hydrogel culture systems with on-demand cell separation

Authors

  • Sharon K. Hamilton,

    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
    2. Department of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA
    3. Department of Chemistry, Auburn University, Chemistry Building, Auburn, AL, USA
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  • Nathaniel C. Bloodworth,

    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
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  • Christopher S. Massad,

    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
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  • Taymour M. Hammoudi,

    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
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  • Shalu Suri,

    1. Department of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA
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  • Peter J. Yang,

    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
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  • Dr. Hang Lu,

    Corresponding author
    1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
    • Department of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Drive NW, Atlanta, GA 30332, USA
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  • Dr. Johnna S. Temenoff

    Corresponding author
    1. Department of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA
    • W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 313 Ferst Drive, Room 2112, Atlanta, GA 30332, USA
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Abstract

Recently there has been an increased interest in the effects of paracrine signaling between groups of cells, particularly in the context of better understanding how stem cells contribute to tissue repair. Most current 3D co-culture methods lack the ability to effectively separate two cell populations after the culture period, which is important for simultaneously analyzing the reciprocal effects of each cell type on the other. Here, we detail the development of a 3D hydrogel co-culture system that allows us to culture different cell types for up to 7 days and subsequently separate and isolate the different cell populations using enzyme-sensitive glues. Separable 3D co-culture laminates were prepared by laminating PEG-based hydrogels with enzyme-degradable hydrogel adhesives. Encapsulated cell populations exhibited good segregation with well-defined interfaces. Furthermore, constructs can be separated on-demand upon addition of the appropriate enzyme, while cell viability remains high throughout the culture period, even after laminate separation. This platform offers great potential for a variety of basic cell signaling studies as the incorporation of an enzyme-sensitive adhesive interface allows the on-demand separation of individual cell populations for immediate analysis or further culture to examine persistence of co-culture effects and paracrine signaling on cell populations.

See accompanying commentary by Danielle R. Bogdanowicz and Helen H. Lu DOI: 10.1002/biot.201300054

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