TGF-β1- and CCN2-Stimulated Sirius Red Assay for Collagen Accumulation in Cultured Cells.

Methods Mol Biol

Department of Molecular and Cell Biology, Henry M. Goldman School of Dental Medicine, Boston University, 700 Albany Street, W-201, Boston, MA, 02118, USA.

Published: January 2018

A simple method for the determination of relative levels of insoluble collagen accumulation in fibroblast cultures is presented. Confluent cell cultures are provided with sodium ascorbate which is then permissive for collagen deposition. At intervals, cultures are fixed and stained successively with sirius red and then crystal Violet to, respectively, assess for relative changes in collagen accumulation in response to factors such as TGF-β1 or matricellular CCN2 and changes in DNA content as an index of changes in cell density.

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http://dx.doi.org/10.1007/978-1-4939-6430-7_39DOI Listing

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