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

Research output: Contribution to journalArticle

Authors

  • K V Greco
  • L Rattazzi
  • G Nalesso
  • N Moradi-Bidhendi
  • A R Moore
  • M B Goldring
  • F Dell'Accio
  • M Perretti

Colleges, School and Institutes

External organisations

  • William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, United Kingdom. k.v.greco@qmul.ac.uk

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.

Details

Original languageEnglish
Pages (from-to)1919-29
Number of pages11
JournalBiochemical Pharmacology
Volume82
Issue number12
Publication statusPublished - 15 Dec 2011

Keywords

  • Alcian Blue, Anti-Inflammatory Agents, Cartilage, Articular, Cell Line, Cell Proliferation, Chondrocytes, Gene Expression Regulation, Humans, Interleukin-1beta, Naproxen, Prednisolone, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't