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Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale

The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve hig...

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Autores principales: Gautrot, Julien E., Trappmann, Britta, Oceguera-Yanez, Fabian, Connelly, John, He, Ximin, Watt, Fiona M., Huck, Wilhelm T.S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712172/
https://www.ncbi.nlm.nih.gov/pubmed/20347135
http://dx.doi.org/10.1016/j.biomaterials.2010.02.066
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author Gautrot, Julien E.
Trappmann, Britta
Oceguera-Yanez, Fabian
Connelly, John
He, Ximin
Watt, Fiona M.
Huck, Wilhelm T.S.
author_facet Gautrot, Julien E.
Trappmann, Britta
Oceguera-Yanez, Fabian
Connelly, John
He, Ximin
Watt, Fiona M.
Huck, Wilhelm T.S.
author_sort Gautrot, Julien E.
collection PubMed
description The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve high fidelity patterning of single cells. These coatings can be patterned by soft lithography on large areas (a microscope slide) and scale (substrates were typically prepared in batches of 200). The present protocol relies on the adsorption of extra-cellular matrix (ECM) proteins on unprotected areas using simple incubation and washing steps. The stability of POEGMA brushes, as examined via ellipsometry and SPR, is found to be excellent, both during storage and cell culture. The impact of substrate treatment, brush thickness and incubation protocol on ECM deposition, both for ultra-thin gold and glass substrates, is investigated via fluorescence microscopy and AFM. Optimised conditions result in high quality ECM patterns at the micron scale, even on glass substrates, that are suitable for controlling cell spreading and polarisation. These patterns are compatible with state-of-the-art technologies (fluorescence microscopy, FRET) used for live cell imaging. This technology, combined with single cell analysis methods, provides a platform for exploring the mechanisms that regulate cell behaviour.
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spelling pubmed-37121722013-07-17 Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale Gautrot, Julien E. Trappmann, Britta Oceguera-Yanez, Fabian Connelly, John He, Ximin Watt, Fiona M. Huck, Wilhelm T.S. Biomaterials Article The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve high fidelity patterning of single cells. These coatings can be patterned by soft lithography on large areas (a microscope slide) and scale (substrates were typically prepared in batches of 200). The present protocol relies on the adsorption of extra-cellular matrix (ECM) proteins on unprotected areas using simple incubation and washing steps. The stability of POEGMA brushes, as examined via ellipsometry and SPR, is found to be excellent, both during storage and cell culture. The impact of substrate treatment, brush thickness and incubation protocol on ECM deposition, both for ultra-thin gold and glass substrates, is investigated via fluorescence microscopy and AFM. Optimised conditions result in high quality ECM patterns at the micron scale, even on glass substrates, that are suitable for controlling cell spreading and polarisation. These patterns are compatible with state-of-the-art technologies (fluorescence microscopy, FRET) used for live cell imaging. This technology, combined with single cell analysis methods, provides a platform for exploring the mechanisms that regulate cell behaviour. Elsevier Science 2010-06 /pmc/articles/PMC3712172/ /pubmed/20347135 http://dx.doi.org/10.1016/j.biomaterials.2010.02.066 Text en © 2010 Elsevier Ltd. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Article
Gautrot, Julien E.
Trappmann, Britta
Oceguera-Yanez, Fabian
Connelly, John
He, Ximin
Watt, Fiona M.
Huck, Wilhelm T.S.
Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title_full Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title_fullStr Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title_full_unstemmed Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title_short Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
title_sort exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712172/
https://www.ncbi.nlm.nih.gov/pubmed/20347135
http://dx.doi.org/10.1016/j.biomaterials.2010.02.066
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