Development of Serum-Free Media for Primary Culture of Human Articular Chondrocytes

  • CHOI YONG SOO (Department of Biological Engineering, Inha University) ;
  • LIM SANG MIN (Department of Biological Engineering, Inha University) ;
  • LEE CHANG WOO (Good Shepherd Hospital) ;
  • KIM DONG-IL (Department of Biological Engineering, Inha University)
  • Published : 2005.12.01

Abstract

Human articular chondrocytes (HAC) were cultivated as a monolayer in a serum-free medium for primary culture (SFM-P). An optimized SFM-P provides $95\%$ proliferation rate of that obtainable from primary and secondary chondrocyte cultures grown in a control medium with serum. The gradual decrease in the amounts of synthesized glycosaminoglycan and type II collagen was improved by coating the culture dishes with type IV collagen and fibronectin. A significant improvement in the expression of type II collagen and aggrecan mRNA could be achieved. In addition, the monolayer cultures showed better synthesis of the extracellular matrices than alginate-bead cultures in SFM-P.

Keywords

References

  1. Adolphe M., B. Froger, X. Ronot, M. T. Corvol, and N. Forest. 1984. Cell multiplication and type II collagen production by rabbit articular chondrocytes cultivated in a defined medium. Exp. Cell Res. 155: 527-536 https://doi.org/10.1016/0014-4827(84)90212-X
  2. Benya, P. D. and J. D. Shaffer. 1982. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30: 215-224 https://doi.org/10.1016/0092-8674(82)90027-7
  3. Biggers, J. D., R. B. L. Gwatkin, and S. Heyner. 1961. Growth of embryonic avian and mammalian tibiae on a relatively simple chemically defined medium. Exp. Cell Res. 25: 41-58 https://doi.org/10.1016/0014-4827(61)90305-6
  4. Brittberg, M., T. Tallheden, E. Sjögren-Jansson, A. Lindahl, and L. Peterson. 2001. Autologous chondrocytes used for articular cartilage repair. Clin. Orthop. Rel. Res. 391S: S337-S348
  5. Choi, Y. C., G. M. Morris, F. S. Lee, and L. Sokoloff. 1980. The effect of serum on monolayer cell culture of mammalian articular chondrocytes. Connect. Tissue Res. 7: 105-112 https://doi.org/10.3109/03008208009152295
  6. Grigolo, B., L. Roseti, S. Neri, P. Gobbi, P. Jensen, E. O. Major, and A. Facchini. 2002. Human articular chondrocytes immortalized by HPV-16 E6 and E7 genes: Maintenance of differentiated phenotype under defined culture conditions. Osteoarthritis Cartilage 10: 879-889 https://doi.org/10.1053/joca.2002.0836
  7. Huckle, J., G. Dootson, N. Medcalf, S. McTaggart, E. Wright, A. Carter, R. Schreiber, B. Kirby, N. Dunkelman, S. Stevenson, S. Riley, T. Davisson, and A. Ratcliffe. 2003. Differentiated chondrocytes for cartilage tissue engineering, pp. 175-186. In G. Bock and J. Goode (eds.), Tissue Engineering of Cartilage and Bone. John Wiley & Sons, West Sussex
  8. Huh, Y. H., S. H. Kim, S. J. Kim, and J. S. Chun. 2003. Differentiation status-dependent regulation of cyclooxygenase- 2 expression and prostaglandin $E_2$ production by epidermal growth factor via mitogen-activated protein kinase in articular chondrocytes. J. Biol. Chem. 278: 9691-9697 https://doi.org/10.1074/jbc.M211360200
  9. Jakob, M., O. Demarteau, D. Schafer, B. Hintermann, W. Dick, M. Heberer, and I. Martin. 2001. Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro. J. Cell Biochem. 81: 368-377 https://doi.org/10.1002/1097-4644(20010501)81:2<368::AID-JCB1051>3.0.CO;2-J
  10. Kim, B.-S., S. I. Jeong, S.-W. Cho, J. Nikolovski, D. J. Mooney, S. H. Lee, J. Jeon, T. W. Kim, S. H. Lim, Y. S. Hong, C. Y. Choi, Y. M. Lee, S. H. Kim, and Y. H. Kim. 2004. Tissue engineering of smooth muscle under a mechanically dynamic condition. J. Microbiol. Biotechnol. 13: 841-845
  11. Kim, B. S., S. P. Yoo, and H. W. Park. 2004. Tissue engineering of cartilage with chondroctyes cultured in a chemically-defined, serum-free medium. Biotechnol. Lett. 26: 709-712 https://doi.org/10.1023/B:BILE.0000024093.94151.d8
  12. Kim, D.-I., H.-J. Park, H.-S. Eo, S.-W. Suh, J.-H. Hong, M.-J. Lee, J.-S. Kim, I.-S. Jang, and B.-S. Kim. 2004. Comparative study of seeding and culture methods to vascular smooth muscle cells on biodegradable scaffold. J. Microbiol. Biotechnol. 14: 707-714
  13. Kim, N., Y. Lee, H. Kim, J. Choi, J. Kim, K.-H. Chang, J. H. Kim, and H.-J. Kim. 2004. Enhancement of erythropoietin production from Chinese hamster ovary (CHO) cells by introduction of the urea cycle enzymes, carbamoyl phosphate synthetase I and ornithine transcarbamylase. J. Microbiol. Biotechnol. 14: 844-851
  14. Kim, N. S., K. H. Chang, B. S. Chung, S. H. Kim, J. H. Kim, and G. M. Lee. 2003. Characterization of humanized antibody produced by apoptosis-resistant CHO cells under sodium butyrate-induced condition. J. Microbiol. Biotechnol. 13: 926-936
  15. Kwon, M. S., T. Dojima, and E. Y. Park. 2003. Comparative characterization of growth and recombinant protein production among three insect cell lines with four kinds of serum free media. Biotechnol. Bioprocess Eng. 8: 142-146 https://doi.org/10.1007/BF02940270
  16. Lu, L., X. Zhu, R. G. Valenzuela, B. L. Currier, and M. J. Yaszemski. 2001. Biodegradable polymer scaffolds for cartilage tissue engineering. Clin. Orthop. Rel. Res. 391S: S251-S270
  17. Prins, A. P. A, J. M. Lipman, and L. Sokoloff. 1982. Effect of purified growth factors on rabbit articular chondrocytes in monolayer culture. Arthritis Rheum. 25: 1217-1227 https://doi.org/10.1002/art.1780251011
  18. Ribault, D., A. M. Khatib, A. Panasyuk, A. Barbara, Z. Bouizar, and R. D. Mitrovic. 1997. Mitogenic and metabolic actions of epidermal growth factor on rat articular chondrocytes: Modulation by fetal calf serum, transforming growth factor-b, and tyrphostin. Arch. Biochem. Biophys. 337: 149-158 https://doi.org/10.1006/abbi.1996.9767
  19. Yashveer, S. 2003. Photosynthetic activity, and lipid and hydrocarbon production by alginate-immobilized cells of Botryococcus in relation to growth phase. J. Microbiol. Biotechnol. 13: 687-691