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3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation
Published on: October 7, 2015
Tissue engineered cartilage using human articular chondrocytes immortalized by HPV-16 E6 and E7 genes
Wei-Hung Chen1, Wen-Fu Lai, Win-Ping Deng
1Graduate Institute of Biomedical Materials, Taipei Medical University, Taipei, Taiwan, Republic of China.
Journal of Biomedical Materials Research. Part A
|November 10, 2005
Summary
Human primary chondrocytes were immortalized using the human papillomavirus type 16 E6/E7 genes, creating a stable cell line for cartilage engineering. This novel cell line preserves cartilage-specific markers for in vitro and in vivo applications.
Area of Science:
- Biotechnology
- Cell Biology
- Tissue Engineering
Background:
- Primary chondrocytes are crucial for studying cartilage degeneration in tissue-engineered cartilage.
- Limitations include short in vitro lifespan and rapid collagen phenotype shifts in primary chondrocytes.
Purpose of the Study:
- To establish an immortalized human chondrocyte cell line for cartilage engineering.
- To assess the preservation of cartilage-specific differentiation markers in the immortalized cell line.
Main Methods:
- Transduction of human primary chondrocytes with a high dosage of retroviral vector LXSN16E6E7.
- Propagation of the resulting cell line (hPi) beyond 100 passages in defined medium.
- Evaluation of cartilage-specific markers (type II collagen, aggrecan, GAG) and tumorigenicity in NOD-SCID mice.
Main Results:
- The immortalized hPi cell line proliferated extensively (over 100 passages) while maintaining key chondrocyte characteristics.
- hPi cells expressed type II collagen and aggrecan mRNA and protein, and showed GAG staining and lacunae formation.
- Tumorigenicity assays confirmed neoplastic transformation of hPi cells.
Conclusions:
- Immortalization of human primary chondrocytes using HPV-16 E6/E7 is feasible, yielding a stable cell line.
- The established hPi cell line retains stable cartilage-specific differentiation markers.
- This novel chondrocyte cell line offers a promising model for in vitro and in vivo cartilage engineering and repair research.
