Maintenance of pluripotency in human embryonic stem cells cultured on a synthetic substrate in conditioned medium

Authors

  • Magdalena M. Mahlstedt,

    1. School of Pharmacy, University of Nottingham, Nottingham, UK
    2. School of Physics and Astronomy, University of Nottingham, Nottingham, UK
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  • David Anderson,

    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling, School of Clinical Sciences, University of Nottingham, Nottingham NG7 2RD, UK; telephone: +44-0-115-82-31236; fax: +44-0-115-31230
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  • James S. Sharp,

    1. School of Physics and Astronomy, University of Nottingham, Nottingham, UK
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  • Roger McGilvray,

    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling, School of Clinical Sciences, University of Nottingham, Nottingham NG7 2RD, UK; telephone: +44-0-115-82-31236; fax: +44-0-115-31230
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  • Maria D. Barbadillo Muñoz,

    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling, School of Clinical Sciences, University of Nottingham, Nottingham NG7 2RD, UK; telephone: +44-0-115-82-31236; fax: +44-0-115-31230
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  • Lee D. Buttery,

    1. School of Pharmacy, University of Nottingham, Nottingham, UK
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  • Morgan R. Alexander,

    1. School of Pharmacy, University of Nottingham, Nottingham, UK
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  • Felicity R.A.J. Rose,

    1. School of Pharmacy, University of Nottingham, Nottingham, UK
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  • Chris Denning

    Corresponding author
    1. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling, School of Clinical Sciences, University of Nottingham, Nottingham NG7 2RD, UK; telephone: +44-0-115-82-31236; fax: +44-0-115-31230
    • Wolfson Centre for Stem Cells, Tissue Engineering & Modelling, School of Clinical Sciences, University of Nottingham, Nottingham NG7 2RD, UK; telephone: +44-0-115-82-31236; fax: +44-0-115-31230.
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Abstract

Realizing the potential clinical and industrial applications of human embryonic stem cells (hESCs) is limited by the need for costly, labile, or undefined growth substrates. Here we demonstrate that trypsin passaging of the hESC lines, HUES7 and NOTT1, on oxygen plasma etched tissue culture polystyrene (PE-TCPS) in conditioned medium is compatible with pluripotency. This synthetic culture surface is stable at room temperature for at least a year and is readily prepared by placing polystyrene substrates in a radio frequency oxygen plasma generator for 5 min. Modification of the polystyrene surface chemistry by plasma etching was confirmed by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), which identified elemental and molecular changes as a result of the treatment. Pluripotency of hESCs cultured on PE-TCPS was gauged by consistent proliferation during serial passage, expression of stem cell markers (OCT4, TRA1-60, and SSEA-4), stable karyotype and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Generation of cost-effective, easy-to-handle synthetic, defined, stable surfaces for hESC culture will expedite stem cell use in biomedical applications. Biotechnol. Bioeng. 2010;105: 130–140. © 2009 Wiley Periodicals, Inc.

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