Fabrication and biological evaluation of uniform extracellular matrix coatings on discontinuous photolithography generated micropallet arrays

J Biomed Mater Res A. 2010 Nov;95(2):401-12. doi: 10.1002/jbm.a.32854.

Abstract

The recent identification of rare cell populations within tissues that are associated with specific biological behaviors, for example, progenitor cells, has illuminated a limitation of current technologies to study such adherent cells directly from primary tissues. The micropallet array is a recently developed technology designed to address this limitation by virtue of its capacity to isolate and recover single adherent cells on individual micropallets. The capacity to apply this technology to primary tissues and cells with restricted growth characteristics, particularly adhesion requirements, is critically dependent on the capacity to generate functional extracellular matrix (ECM) coatings. The discontinuous nature of the micropallet array surface provides specific constraints on the processes for generating the desired ECM coatings that are necessary to achieve the full functional capacity of the micropallet array. We have developed strategies, reported herein, to generate functional coatings with various ECM protein components: fibronectin, EHS tumor basement membrane extract, collagen, and laminin-5; confirmed by evaluation for rapid cellular adherence of four dissimilar cell types: fibroblast, breast epithelial, pancreatic epithelial, and myeloma. These findings are important for the dissemination and expanded use of micropallet arrays and similar microtechnologies requiring the integrated use of ECM protein coatings to promote cellular adherence.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Adhesion Molecules / metabolism
  • Cell Adhesion*
  • Cell Line
  • Coated Materials, Biocompatible / chemistry*
  • Collagen / metabolism
  • Extracellular Matrix / chemistry*
  • Extracellular Matrix Proteins / chemistry
  • Extracellular Matrix Proteins / metabolism
  • Fibronectins / metabolism
  • Humans
  • Kalinin
  • Materials Testing
  • Microarray Analysis* / instrumentation
  • Microarray Analysis* / methods
  • Photochemistry* / instrumentation
  • Photochemistry* / methods
  • Surface Properties

Substances

  • Cell Adhesion Molecules
  • Coated Materials, Biocompatible
  • Extracellular Matrix Proteins
  • Fibronectins
  • Collagen