Protocol

Developmental Biology Protocols

Volume 137 of the series Methods in Molecular Biology™ pp 359-375

Embryonic Limb Mesenchyme Micromass Culture as an In Vitro Model for Chondrogenesis and Cartilage Maturation

  • Anthony M. De LiseAffiliated withDepartment of Orthopaedic Surgery, Thomas Jefferson University
  • , Emanuela StringaAffiliated withDepartment of Orthopaedic Surgery, Thomas Jefferson University
  • , Wendy A. WoodwardAffiliated withDepartment of Orthopaedic Surgery, Thomas Jefferson University
  • , Maria Alice MelloAffiliated withDepartment of Orthopaedic Surgery, Thomas Jefferson University
  • , Rocky S. TuanAffiliated withDepartment of Orthopaedic Surgery, Thomas Jefferson University

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Abstract

In vitro techniques for the study of chondrogenic differentiation of embryonic limb mesenchymal cells have been available for some time. Early methods require highdensity confluent monolayer cell cultures (1,2). The micromass culture method developed by Ahrens et al. (3) represented a convenient system for the observations and analysis of the differentiative processes and phenomena analogous to those exhibited by the limb cartilage anlagen in situ. In these cultures, limb mesenchymal cells first undergo condensation giving rise to aggregates that later become cartilage nodules (3,4), thereby mimicking the differentiative phenomena occurring during embryonic limb development in vivo, i.e., mesenchymal condensation preceding cartilage differentiation (59). The micromass limb mesenchymal culture system has gained great popularity for the analysis of the regulatory steps and differentiative processes that result in the condensation of the mesenchyme and the formation and maturation of the cartilage anlagen.