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

BMC Biotechnology
Kurt J LangenbachAnne L Plant

Abstract

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. 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. 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.

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Citations

Oct 5, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Ryan R HansenKeith B Neeves
Nov 2, 2012·Proceedings of the National Academy of Sciences of the United States of America·Daniel R SisanAnne L Plant
Dec 11, 2008·Annual Review of Entomology·Florian P Schiestl, Philipp M Schlüter
Dec 13, 2006·Biophysical Journal·Dennis P McDanielAnne L Plant
Nov 26, 2008·Biochimica Et Biophysica Acta·Anne L PlantJohn T Elliott
May 24, 2012·Biotechnology Progress·Annalaura ManciaAnne L Plant
Sep 11, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Michael HalterJohn T Elliott
Feb 21, 2008·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Franck AmyotAmir Gandjbakhche
Nov 7, 2012·Journal of Microscopy·J ChalfounM Brady
Nov 3, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Michael HalterJohn T Elliott
Jan 20, 2021·ACS Biomaterials Science & Engineering·Vittorio FerraraBruno Pignataro
Dec 11, 2008·Journal of Theoretical Biology·Michael HalterAnne L Plant

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Methods Mentioned

BETA
fluorescence microscopy
light microscopy
flow cytometry
transfection

Software Mentioned

Sigma Stat
ISee

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