Exploiting the Superior Protein Resistance of Polymer Brushes to Control Single Cell Adhesion and Polarisation at the Micron Scale

Biomaterials. 2010 Jun;31(18):5030-41. doi: 10.1016/j.biomaterials.2010.02.066. Epub 2010 Mar 26.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Adsorption
  • Animals
  • Blood Proteins / metabolism
  • Cell Adhesion*
  • Cell Polarity*
  • Cells, Cultured
  • Coated Materials, Biocompatible / chemistry
  • Coated Materials, Biocompatible / metabolism*
  • Extracellular Matrix Proteins / chemistry
  • Glass / chemistry
  • Gold / chemistry
  • Humans
  • Keratinocytes / cytology
  • Methacrylates / chemistry
  • Methacrylates / metabolism*
  • Mice
  • Polyethylene Glycols / chemistry
  • Polyethylene Glycols / metabolism*
  • Polymethacrylic Acids
  • Protein Binding

Substances

  • Blood Proteins
  • Coated Materials, Biocompatible
  • Extracellular Matrix Proteins
  • Methacrylates
  • Polymethacrylic Acids
  • poly(poly(ethylene glycol)methacrylate)
  • Polyethylene Glycols
  • Gold