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High density micromass cultures of a human chondrocyte cell line: A reliable assay system to reveal the modulatory functions of pharmacological agents

Greco, KV, Iqbal, AJ, Rattazzi, L, Nalesso, G, Moradi-Bidhendi, N, Moore, AR, Goldring, MB, Dell’Accio, F and Perretti, M (2011) High density micromass cultures of a human chondrocyte cell line: A reliable assay system to reveal the modulatory functions of pharmacological agents Biochemical Pharmacology, 82 (12). pp. 1919-1929.

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Abstract

Osteoarthritis is a highly prevalent and disabling disease for which we do not have a cure. The identification of suitable molecular targets is hindered by the lack of standardized, reproducible and convenient screening assays. Following extensive comparisons of a number of chondrocytic cell lines, culture conditions, and readouts, we have optimized an assay utilizing C-28/I2, a chondrocytic cell line cultured in high-density micromasses. Utilizing molecules with known effects on cartilage (e.g. IL-1β, TGFβ1, BMP-2), we have exploited this improved protocol to (i) evoke responses characteristic of primary chondrocytes; (ii) assess the pharmacodynamics of gene over-expression using non-viral expression vectors; (iii) establish the response profiles of known pharmacological treatments; and (iv) investigate their mechanisms of action. These data indicate that we have established a medium-throughput methodology for studying chondrocyte-specific cellular and molecular responses (from gene expression to rapid quantitative measurement of sulfated glycosaminoglycans by Alcian blue staining) that may enable the discovery of novel therapeutics for pharmacological modulation of chondrocyte activation in osteoarthritis.

Item Type: Article
Subjects : Veterinary Medicine
Authors :
NameEmailORCID
Greco, KVUNSPECIFIEDUNSPECIFIED
Iqbal, AJUNSPECIFIEDUNSPECIFIED
Rattazzi, LUNSPECIFIEDUNSPECIFIED
Nalesso, Gg.nalesso@surrey.ac.ukUNSPECIFIED
Moradi-Bidhendi, NUNSPECIFIEDUNSPECIFIED
Moore, ARUNSPECIFIEDUNSPECIFIED
Goldring, MBUNSPECIFIEDUNSPECIFIED
Dell’Accio, FUNSPECIFIEDUNSPECIFIED
Perretti, MUNSPECIFIEDUNSPECIFIED
Date : December 2011
Identification Number : https://doi.org/10.1016/j.bcp.2011.09.009
Copyright Disclaimer : 2011 Elsevier Inc. All rights reserved.
Uncontrolled Keywords : IL-1β, TGFβ1, Micromasses, Anti-inflammatory drugs, Naproxen, Prednisolone
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 10:48
Last Modified : 18 May 2017 12:44
URI: http://epubs.surrey.ac.uk/id/eprint/829285

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