Human chondrocyte cultures as models of cartilage-specific gene regulation

Miguel Otero, Marta Favero, Cecilia Dragomir, Karim El Hachem, Ko Hashimoto, Darren A. Plumb, Mary B. Goldring

Research output: Chapter in Book/Report/Conference proceedingChapter

27 Citations (Scopus)

Abstract

The human adult articular chondrocyte is a unique cell type that has reached a fully differentiated state as an end point of development. Within the cartilage matrix, chondrocytes are normally quiescent and maintain the matrix constituents in a low-turnover state of equilibrium. Isolated chondrocytes in culture have provided useful models to study cellular responses to alterations in the environment such as those occurring in different forms of arthritis. However, expansion of primary chondrocytes in monolayer culture results in the loss of phenotype, particularly if high cell density is not maintained. This chapter describes strategies for maintaining or restoring differentiated phenotype by culture in suspension, gels, or scaffolds. Techniques for assessing phenotype involving primarily the analysis of synthesis of cartilage-specific matrix proteins as well as the corresponding mRNAs are also described. Approaches for studying gene regulation, including transfection of promoter-driven reporter genes with expression vectors for transcriptional and signaling regulators, chromatin immunoprecipitation, and DNA methylation are also described.

Original languageEnglish
Title of host publicationHuman Cell Culture Protocols
EditorsRagai Mitry, Robin Hughes
Pages301-336
Number of pages36
DOIs
Publication statusPublished - 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume806
ISSN (Print)1064-3745

Keywords

  • Adenovirus-mediated expression
  • Agarose
  • Aggrecan
  • Alginate
  • Chondrocyte
  • Chromatin immunoprecipitation assay
  • Collagen scaffolds
  • Luciferase reporter plasmids
  • Monolayer culture
  • PolyHEMA
  • Suspension culture
  • Three-dimensional scaffolds
  • Transfections
  • Type II collagen

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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