Establishment and characterization of chondrocyte cell lines from the costal cartilage of SV40 large T antigen transgenic mice

Complete understanding of the physiology and pathology of the cartilage is essential to establish treatments for a variety of cartilage disorders and defects such as rheumatoid arthritis, congenital malformations, and tumors of cartilage. Although synthetic materials have been used in many cases, th ey possess inherent problems including wear of the materials and low mechan ical strength. Autograft has been considered very effective to overcome the se problems. However, the limitation of the transplant volume is a major pr oblem in autograft to be overcome. The costal cartilage is the most serious candidate for donor site transplantation, since it is the largest permanen t hyaline cartilage in the body. To investigate the possibility using the c ostal cartilage as a transplant source, we have established and characteriz ed three mouse chondrocyte cell lines (MCC-2, MCC-5, and MCC-35) derived fr om the costal cartilage of 8-week-old male SV40 large T-antigen transgenic mice. At confluence, all the cell lines formed nodules that could be positi vely stained with alcian blue (pH 2.5). The size of nodules gradually incre ased during culturing time. After 2 and 6 weeks of culture, RT-PCR analysis demonstrated that all three cell lines expressed mRNA from the cartilage-s pecific genes for type II collagen, type XI collagen, aggrecan, and link pr otein. Furthermore, type X collagen expression was detected in MCC-5 and MC C-35 but not in MCC-2. Any phenotypic changes were not observed over 31 cel l divisions. Immunocytochemistry showed further that MCC-2, MCC-5, and MCC- 35 produced cartilage-specific proteins type II collagen and type XI collag en, while in addition MCC-5 and MCC-35 produced type X collagen. Treatment with 1 alpha. 25-dihydroxyvitamin D-3 inhibited cell proliferation and diff erentiation of the three cell lines in a dose-dependent manner. These pheno typic characteristics have been found consistent with chondrocyte cell line s established from cartilage tissues other than costal cartilage. In conclu sion, costal cartilage shows phenotypic similarities to other cartilages, i .e., articular cartilage and embryonic limbs, suggesting that costal cartil age may be very useful as the donor transplantation site for the treatment of cartilage disorders. Furthermore, the cell lines established in this stu dy are also beneficial in basic research of cartilage physiology and pathol ogy. J. Cell. Biochem. 81:571-582, 2001. (C) 2001 Wiley-Liss. Inc.