Tissue engineered cartilage using human articular chondrocytes immortalized by HPV-16 E6 and E7 genes

Wei-Hung Chen, Wen-FuThomas Lai, Win Ping Deng, Wen K. Yang, Wen Cheng Lo, Cheng Chia Wu, Den Mei Yang, Ming-Tang Lai, Che Tong Lin, Tsou Wen Lin, Charng Bin Yang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Chondrocytes are useful as a cell culture system for studying arthritic degeneration in tissue engineered cartilage. However, primary chondrocytes have short in vitro lifespan and rapid shift of collagen phenotype. In this study, we used a high dosage of retroviral vector LXSN16E6E7 to transduce human primary chondrocytes and obtained an actively proliferating cell line, designated hPi, which expresses HPV-16 E6/E7 mRNA in early passages. Parental primary chondrocytes cease to grow after five passages, whereas hPi could be propagated beyond 100 passages without requiring additional cell elements in defined medium. After 48 passages, hPi can also give many profiles similar to those of parental primary chondrocyte, including type II collagen in mRNA and protein level, aggrecan in mRNA level, lacunae in type I collagen matrices, and morphology with GAG-specific Alcian blue staining. hPi has shown neoplastic transformation, as examined by NOD-SCID mice tumorgenicity assays for 3 months. Our results indicated that human primary chondrocytes could be immortalized by transduction with HPV-16 E6/E7, preserving stable cartilage-specific differentiation markers. The established chondrocyte cell line could provide a novel model to engineer cartilage in vitro and in vivo for cartilage repair research and clinical application.

Original languageEnglish
Pages (from-to)512-520
Number of pages9
JournalJournal of Biomedical Materials Research - Part A
Volume76
Issue number3
DOIs
Publication statusPublished - Mar 1 2006

Keywords

  • Articular cartilage
  • Cell differentiation
  • Extracellular matrix
  • HPV-16 E6/E7
  • Immortalization

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

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