ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes

Emi Matsumoto, Takayuki Furumatsu, Tomoko Kanazawa, Masanori Tamura, Toshihumi Ozaki

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Chondrocytes lose their chondrocytic phenotypes in vitro. The Rho family GTPase ROCK, involved in organizing the actin cytoskeleton, modulates the differentiation status of chondrocytic cells. However, the optimum method to prepare a large number of un-dedifferentiated chondrocytes is still unclear. In this study, we investigated the effect of ROCK inhibitor (ROCKi) on the chondrogenic property of monolayer-cultured articular chondrocytes. Human articular chondrocytes were subcultured in the presence or absence of ROCKi (Y-27632). The expression of chondrocytic marker genes such as SOX9 and COL2A1 was assessed by quantitative real-time PCR analysis. Cellular morphology and viability were evaluated. Chondrogenic redifferentiation potential was examined by a pellet culture procedure. The expression level of SOX9 and COL2A1 was higher in ROCKi-treated chondrocytes than in untreated cells. Chondrocyte morphology varied from a spreading form to a round shape in a ROCKi-dependent manner. In addition, ROCKi treatment stimulated the proliferation of chondrocytes. The deposition of safranin O-stained proteoglycans and type II collagen was highly detected in chondrogenic pellets derived from ROCKi-pretreated chondrocytes. Our results suggest that ROCKi prevents the dedifferentiation of monolayer-cultured chondrocytes, and may be a useful reagent to maintain chondrocytic phenotypes in vitro for chondrocyte-based regeneration therapy.

Original languageEnglish
Pages (from-to)124-129
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume420
Issue number1
DOIs
Publication statusPublished - Mar 30 2012

Fingerprint

Chondrocytes
Monolayers
Joints
Collagen Type II
GTP Phosphohydrolases
Proteoglycans
Actins
Genes
Phenotype
rho GTP-Binding Proteins
Actin Cytoskeleton
Real-Time Polymerase Chain Reaction
Regeneration
safranine T
Y 27632

Keywords

  • Chondrocyte
  • Dedifferentiation
  • Redifferentiation
  • ROCK inhibitor
  • SOX9
  • Type II collagen

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes. / Matsumoto, Emi; Furumatsu, Takayuki; Kanazawa, Tomoko; Tamura, Masanori; Ozaki, Toshihumi.

In: Biochemical and Biophysical Research Communications, Vol. 420, No. 1, 30.03.2012, p. 124-129.

Research output: Contribution to journalArticle

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