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Analysis of extracellular matrix proteins produced by cultured cells
Institute of Cytology RAS, St. Petersburg, Russian Federation.
Institute of Cytology RAS, St. Petersburg, Russian Federation.
Institute of Cytology RAS, St. Petersburg, Russian Federation.
Linköping University, Department of Clinical and Experimental Medicine, Medical Microbiology. Linköping University, Faculty of Health Sciences.
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2009 (English)In: Cell and Tissue Biology, ISSN 1990-519X, Vol. 3, no 5, 497-502 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
SP MAIK Nauka/Interperiodica , 2009. Vol. 3, no 5, 497-502 p.
Keyword [en]
extracellular matrix, fibroblasts, collagen, fibronectin
National Category
Medical and Health Sciences Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:liu:diva-57559DOI: 10.1134/S1990519X09050137OAI: oai:DiVA.org:liu-57559DiVA: diva2:326550
Note

The original version in Russian:  L.V. Turoverova, M.G. Khotin, N.M. Yudintseva, K.-E. Magnusson, M.I. Blinova, G.P. Pinaev, D.G. Tentler, 2009, published in Tsitologiya, Vol. 51, No. 8, 2009, pp. 691–696. 

Available from: 2010-06-23 Created: 2010-06-23 Last updated: 2017-01-19Bibliographically approved

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Magnusson, Karl-Eric

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