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Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

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Abstract

Cell–cell interaction is an integral part of embryoid body (EB) formation controlling 3D aggregation. Manipulation of embryonic stem (ES) cell interactions could provide control over EB formation. Studies have shown a direct relationship between EB formation and ES cell differentiation. We have previously described a cell surface modification and cross-linking method for influencing cell–cell interaction and formation of multicellular constructs. Here we show further characterisation of this engineered aggregation. We demonstrate that engineering accelerates ES cell aggregation, forming larger, denser and more stable EBs than control samples, with no significant decrease in constituent ES cell viability. However, extended culture ≥5 days reveals significant core necrosis creating a layered EB structure. Accelerated aggregation through engineering circumvents this problem as EB formation time is reduced. We conclude that the proposed engineering method influences initial ES cell-ES cell interactions and EB formation. This methodology could be employed to further our understanding of intrinsic EB properties and their effect on ES cell differentiation.

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Acknowledgements

We would like to thank the Histopathology Department of the Queens Medical Centre, Nottingham for their help with EB sectioning and H&E stains. Thanks also to Dr. Daniel Howard and Dr. Magdalen Self for their advice and guidance. Thanks also to technicians Mrs Christine Grainger-Boultby and Mrs Teresa Marshall for help with equipment training. A final thanks to the BBSRC for funding the research.

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Authors and Affiliations

  1. STEM, Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK

    David Gothard, Kevin M. Shakesheff & Lee D. Buttery

  2. Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven, O&N 1, Herestraat 49 bus 813, 3000, Leuven, Belgium

    Scott J. Roberts

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  1. David Gothard
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  4. Lee D. Buttery
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Correspondence to Lee D. Buttery.

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Gothard, D., Roberts, S.J., Shakesheff, K.M. et al. Controlled embryoid body formation via surface modification and avidin–biotin cross-linking. Cytotechnology 61, 135–144 (2009). https://doi.org/10.1007/s10616-010-9255-3

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  • DOI: https://doi.org/10.1007/s10616-010-9255-3

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