Thin films of Type 1 collagen for cell by cell analysis of morphology and tenascin-C promoter activity

BMC Biotechnol. 2006 Mar 6:6:14. doi: 10.1186/1472-6750-6-14.

Abstract

Background: The use of highly reproducible and spatiallyhomogeneous thin film matrices permits automated microscopy and quantitative determination of the response of hundreds of cells in a population. Using thin films of extracellular matrix proteins, we have quantified, on a cell-by-cell basis, phenotypic parameters of cells on different extracellular matrices. We have quantitatively examined the relationship between fibroblast morphology and activation of the promoter for the extracellular matrix protein tenascin-C using a tenascin-C promoter-based GFP reporter construct.

Results: We find that when considering the average response from the population of cells, cell area correlates with tenascin-C promoter activity as has been previously suggested; however cell-by-cell analysis suggests that cell area and promoter activity are not tightly correlated within individual cells.

Conclusion: This study demonstrates how quantitative cell-by-cell analysis, facilitated by the use of thin films of extracellular matrix proteins, can provide insight into the relationship between phenotypic parameters.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins / metabolism
  • Animals
  • Cell Communication
  • Collagen Type I / chemistry*
  • Cytoskeleton / ultrastructure
  • Extracellular Matrix Proteins / chemistry
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism*
  • Gene Expression Regulation
  • Green Fluorescent Proteins
  • Mice
  • NIH 3T3 Cells
  • Promoter Regions, Genetic / physiology*
  • Tenascin / genetics*

Substances

  • Actins
  • Collagen Type I
  • Extracellular Matrix Proteins
  • Tenascin
  • Green Fluorescent Proteins