Cell and Tissue Biology

, 3:497

Analysis of extracellular matrix proteins produced by cultured cells

Authors

    • Institute of CytologyRussian Academy of Sciences
  • M. G. Khotin
    • Institute of CytologyRussian Academy of Sciences
  • N. M. Yudintseva
    • Institute of CytologyRussian Academy of Sciences
  • K. -E. Magnusson
    • Department of Clinical and Experimental MedicineLinkoping University
  • M. I. Blinova
    • Institute of CytologyRussian Academy of Sciences
  • G. P. Pinaev
    • Institute of CytologyRussian Academy of Sciences
  • D. G. Tentler
    • Institute of CytologyRussian Academy of Sciences
Article

DOI: 10.1134/S1990519X09050137

Cite this article as:
Turoverova, L.V., Khotin, M.G., Yudintseva, N.M. et al. Cell Tiss. Biol. (2009) 3: 497. doi:10.1134/S1990519X09050137

Abstract

The extracellular matrix (ECM) is a highly organized multimolecular structure essential for the vital functions of any organism. Although much of the data of extracellular matrix components has been accumulated, the isolation of an entire set of these proteins remains a complex procedure due to the high content of fibrillar proteins and proteoglycans, which form multidomain, netlike structures. In the study presented, we developed a method for isolating ECM proteins from cell cultures. Human epidermoid carcinoma cells A431 and fibroblasts obtained from normal and scar human skin were used. We showed that EDTA solution removed cells from culture plates without destroying the cell membranes. Subsequent treatment of remaining ECM proteins with acetic acid in order to dissociate collagen fibers significantly improved the fractioning of ECM proteins. The extraction of remaining proteins from the surface of the culture plate was preformed by a buffer developed based on Laemmli probe buffer. Using this method, we isolated ECM proteins synthesized by cultured cells, and the extracted proteins were suitable for future analysis by SDS PAGE and two-dimentional electrophoresis, as well as for identifying individual proteins by mass spectrometry. This study may allow us to compare assortments of ECM proteins isolated from different sources, and elucidate impact of various proteins on structure and property of extracellular matrix of investigated cells.

Key words

extracellular matrix fibroblasts collagen fibronectin

Abbreviations

ECM

extracellular matrix

Copyright information

© Pleiades Publishing, Ltd. 2009