Automated, scalable culture of human embryonic stem cells in feeder-free conditions

Authors

  • Rob J. Thomas,

    Corresponding author
    1. Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44-1509-22-7601; fax: +44-1509-22-7615
    • Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44-1509-22-7601; fax: +44-1509-22-7615.
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  • David Anderson,

    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK, telephone +44-115-82-31236; Fax: +44-115-82-31230
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  • Amit Chandra,

    1. Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44-1509-22-7601; fax: +44-1509-22-7615
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  • Nigel M. Smith,

    1. Department of Cytogenetics, Centre for Medical Genetics, Nottingham City Hospital NHS Trust, Nottingham, UK
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  • Lorraine E. Young,

    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK, telephone +44-115-82-31236; Fax: +44-115-82-31230
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  • David Williams,

    1. Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44-1509-22-7601; fax: +44-1509-22-7615
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  • Chris Denning

    Corresponding author
    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK, telephone +44-115-82-31236; Fax: +44-115-82-31230
    • Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44-1509-22-7601; fax: +44-1509-22-7615.
    Search for more papers by this author

Abstract

Large-scale manufacture of human embryonic stem cells (hESCs) is prerequisite to their widespread use in biomedical applications. However, current hESC culture strategies are labor-intensive and employ highly variable processes, presenting challenges for scaled production and commercial development. Here we demonstrate that passaging of the hESC lines, HUES7, and NOTT1, with trypsin in feeder-free conditions, is compatible with complete automation on the CompacT SelecT, a commercially available and industrially relevant robotic platform. Pluripotency was successfully retained, as evidenced by consistent proliferation during serial passage, expression of stem cell markers (OCT4, NANOG, TRA1-81, and SSEA-4), stable karyotype, and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Automation of hESC culture will expedite cell-use in clinical, scientific, and industrial applications. Biotechnol. Bioeng. 2009;102: 1636–1644. © 2008 Wiley Periodicals, Inc.

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