A thermoresponsive and chemically defined hydrogel for long-term culture of human embryonic stem cells

Rong Zhang, Heidi K. Mjoseng, Marieke A. Hoeve, Nina G. Bauer, Steve Pells, Rut Besseling, Srinivas Velugotla, Guilhem Tourniaire, Ria E. B. Kishen, Yanina Tsenkina, Chris Armit, Cairnan R. E. Duffy, Martina Helfen, Frank Edenhofer, Paul A. de Sousa () and Mark Bradley ()
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Rong Zhang: EaStChem, School of Chemistry, University of Edinburgh
Heidi K. Mjoseng: Scottish Centre for Regenerative Medicine, University of Edinburgh
Marieke A. Hoeve: Scottish Centre for Regenerative Medicine, University of Edinburgh
Nina G. Bauer: Scottish Centre for Regenerative Medicine, University of Edinburgh
Steve Pells: Scottish Centre for Regenerative Medicine, University of Edinburgh
Rut Besseling: Merck Sharp & Dohme
Srinivas Velugotla: School of Engineering and Electronics, Institute for Integrated Micro and Nano Systems, University of Edinburgh
Guilhem Tourniaire: EaStChem, School of Chemistry, University of Edinburgh
Ria E. B. Kishen: Scottish Centre for Regenerative Medicine, University of Edinburgh
Yanina Tsenkina: Scottish Centre for Regenerative Medicine, University of Edinburgh
Chris Armit: Scottish Centre for Regenerative Medicine, University of Edinburgh
Cairnan R. E. Duffy: Scottish Centre for Regenerative Medicine, University of Edinburgh
Martina Helfen: Institute of Reconstructive Neurobiology, Stem Cell Engineering Group, University of Bonn, Life & Brain Center and Hertie Foundation
Frank Edenhofer: Institute of Reconstructive Neurobiology, Stem Cell Engineering Group, University of Bonn, Life & Brain Center and Hertie Foundation
Paul A. de Sousa: Scottish Centre for Regenerative Medicine, University of Edinburgh
Mark Bradley: EaStChem, School of Chemistry, University of Edinburgh

Nature Communications, 2013, vol. 4, issue 1, 1-10

Abstract: Abstract Cultures of human embryonic stem cell typically rely on protein matrices or feeder cells to support attachment and growth, while mechanical, enzymatic or chemical cell dissociation methods are used for cellular passaging. However, these methods are ill defined, thus introducing variability into the system, and may damage cells. They also exert selective pressures favouring cell aneuploidy and loss of differentiation potential. Here we report the identification of a family of chemically defined thermoresponsive synthetic hydrogels based on 2-(diethylamino)ethyl acrylate, which support long-term human embryonic stem cell growth and pluripotency over a period of 2–6 months. The hydrogels permitted gentle, reagent-free cell passaging by virtue of transient modulation of the ambient temperature from 37 to 15 °C for 30 min. These chemically defined alternatives to currently used, undefined biological substrates represent a flexible and scalable approach for improving the definition, efficacy and safety of human embryonic stem cell culture systems for research, industrial and clinical applications.

Date: 2013
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DOI: 10.1038/ncomms2341

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