Real-time monitoring and control of soluble signaling factors enables enhanced progenitor cell outputs from human cord blood stem cell cultures

Authors

  • Elizabeth Csaszar,

    1. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada
    2. Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada
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  • Kun Chen,

    1. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada
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  • Julia Caldwell,

    1. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada
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  • Warren Chan,

    1. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada
    2. Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada
    3. Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada
    4. Department of Chemistry, University of Toronto, Toronto, Ontario
    5. Department of Materials Science and Engineering, University of Toronto, Toronto, Ontario
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  • Peter W. Zandstra

    Corresponding author
    1. Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada
    2. Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada
    3. Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada
    4. Heart and Stroke/Richard Lewar Centre of Excellence, University of Toronto, Toronto, Ontario, Canada
    5. McEwen Centre for Regenerative Medicine, University Health Network, Toronto Medical Discovery Tower, Toronto, Ontario, Canada
    • Correspondence to: P.W. Zandstra

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  • Elizabeth Csaszar and Kun Chen contributed equally to this work.

ABSTRACT

Monitoring and control of primary cell cultures is challenging as they are heterogenous and dynamically complex systems. Feedback signaling proteins produced from off-target cell populations can accumulate, inhibiting the production of the desired cell populations. Although culture strategies have been developed to reduce feedback inhibition, they are typically optimized for a narrow range of process parameters and do not allow for a dynamically regulated response. Here we describe the development of a microbead-based process control system for the monitoring and control of endogenously produced signaling factors. This system uses quantum dot barcoded microbeads to assay endogenously produced signaling proteins in the culture media, allowing for the dynamic manipulation of protein concentrations. This monitoring system was incorporated into a fed-batch bioreactor to regulate the accumulation of TGF-β1 in an umbilical cord blood cell expansion system. By maintaining the concentration of TGF-β1 below an upper threshold throughout the culture, we demonstrate enhanced ex vivo expansion of hematopoietic progenitor cells at higher input cell densities and over longer culture periods. This study demonstrates the potential of a fully automated and integrated real-time control strategy in stem cell culture systems, and provides a powerful strategy to achieve highly regulated and intensified in vitro cell manufacturing systems. Biotechnol. Bioeng. 2014;111: 1258–1264. © 2013 The Authors Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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