Chondrocyte differentiation in a rat mesenchymal cell line

Gregory P. Lunstrum, Douglas R. Keene, Nicole B. Weksler, Yoon-Jae Cho, Marcus Cornwall, William A. Horton

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

We used a combination of morphologic and histochemical methods to demonstrate that rat calvaria-derived mesenchymal cells, RCJ 3.1C5.18, in culture progress through the differentiation pathway exhibited by chondrocytes in the endochondral growth plate. The cells were grown either as monolayer or suspension cultures. Subconfluent monolayer cultures did not express markers typical of chondrocyte phenotypes. However, after reaching confluency the cells formed nodules of chondrocytic cells separated by cartilage-appearing matrix and encapsulated by fibroblast-like cells. Suspension culture produced cell aggregates with similar characteristics. Matrix in both the nodules and aggregates stained for collagen Types II and XI and aggrecan, and some cells displayed a distinctive pericellular matrix that stained for Type X collagen. Mineralization was evident in older cultures. By electron microscopy, most cells in the aggregates appeared as typical chondrocytes. However, some larger cells were surrounded by a 'mat' of matrix comprised of hexagonal arrays of dense nodules interconnected by a filamentous network. Immunogold localization confirmed the presence of collagen Type X in this matrix. Analysis of markers of chondrocyte differentiation and terminal differentiation over time showed that these markers were acquired sequentially over 2 weeks of culture. This model system will be useful to study the regulation of various steps in the chondrocyte differentiation pathway.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalJournal of Histochemistry and Cytochemistry
Volume47
Issue number1
DOIs
StatePublished - Jan 1 1999

Fingerprint

Chondrocytes
Cell Line
Collagen Type X
Suspensions
Collagen Type XI
Aggrecans
Growth Plate
Collagen Type II
Differentiation Antigens
Skull
Cartilage
Electron Microscopy
Cell Culture Techniques
Fibroblasts
Phenotype

Keywords

  • Cartilage
  • Cell culture
  • Chondrocyte
  • Differentiation
  • Growth Plate
  • Histochemistry
  • Type X collagen

ASJC Scopus subject areas

  • Anatomy
  • Histology

Cite this

Chondrocyte differentiation in a rat mesenchymal cell line. / Lunstrum, Gregory P.; Keene, Douglas R.; Weksler, Nicole B.; Cho, Yoon-Jae; Cornwall, Marcus; Horton, William A.

In: Journal of Histochemistry and Cytochemistry, Vol. 47, No. 1, 01.01.1999, p. 1-6.

Research output: Contribution to journalArticle

Lunstrum, Gregory P. ; Keene, Douglas R. ; Weksler, Nicole B. ; Cho, Yoon-Jae ; Cornwall, Marcus ; Horton, William A. / Chondrocyte differentiation in a rat mesenchymal cell line. In: Journal of Histochemistry and Cytochemistry. 1999 ; Vol. 47, No. 1. pp. 1-6.
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