Applied Biochemistry and Biotechnology

, Volume 163, Issue 1, pp 1–13 | Cite as

Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media

  • Fabiana R. X. Batista
  • Kátia N. Greco
  • Renato M. Astray
  • Soraia A. C. Jorge
  • Elisabeth F. P. Augusto
  • Carlos A. Pereira
  • Ronaldo Z. Mendonça
  • Ângela M. MoraesEmail author


An animal protein-free medium composed of IPL-41 containing 6 g L−1 yeastolate ultrafiltrate, 10 g L−1 glucose, 2 g L−1 lactose, 5 g L−1 glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2 × 107 cells mL−1 in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL−1 and 638 ng RVGP mL−1, respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL−1 and 322 ng HBsAg mL−1 in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3 × 107 cells mL−1 and produced up to 758 ng RVGP mL−1. In general, glucose was consumed by cells, while lactate and ammonia were produced.


S2 cells Metabolism RVGP HBsAg Animal protein-free medium 



The authors acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, Brazil), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant no. 02/09482-3, São Paulo, Brazil), Instituto de Pesquisas Tecnológicas do Estado de São Paulo (IPT, São Paulo, Brazil) and Instituto Butantan (São Paulo, Brazil) for the financial support. The authors gratefully acknowledge M Aguiar and R Andrade (IPT, São Paulo, Brazil) for the technical assistance. AM Moraes and CA Pereira are recipients of CNPq fellowships.


  1. 1.
    Butler, M. (2006). Cytotechnology, 50, 57–76.CrossRefGoogle Scholar
  2. 2.
    Maiorella, B., Inlow, D., Shauger, A., & Harano, D. (1998). Nature Biotechnology, 6, 1406–1410.Google Scholar
  3. 3.
    Akhnoukh, R., Kretzmer, G., & Schügert, K. (1996). Enzyme and Microbial Technology, 18, 220–228.CrossRefGoogle Scholar
  4. 4.
    Benting, J., Lecat, S., Zacchetti, D., & Simons, K. (2000). Analytical Biochemistry, 278, 59–68.CrossRefGoogle Scholar
  5. 5.
    Taticek, R. A., Choi, C., Phan, S.-E., Palomares, L. A., & Shuler, M. L. (2001). Biotechnology Progress, 17, 676–684.CrossRefGoogle Scholar
  6. 6.
    Ikonomou, L., Bastin, G., Schneider, Y. J., & Agathos, S. N. (2001). In Vitro Cellular & Developmental Biology, Animal, 37, 549–559.CrossRefGoogle Scholar
  7. 7.
    Valle, M. A., Kester, M. B., Burns, A. L., Marx, S. J., Spiegel, A. M., & Shiloach, J. (2001). Cytotechnology, 35, 127–135.CrossRefGoogle Scholar
  8. 8.
    Elias, C., Carpentier, B., Durocher, E., Bisson, Y., Wagner, L., & Kamen, R. (2003). Biotechnology Progress, 19, 90–97.CrossRefGoogle Scholar
  9. 9.
    Shin, H., & Cha, H. J. (2002). Biotechnology Progress, 18, 1187–1194.CrossRefGoogle Scholar
  10. 10.
    Shin, H. S., & Cha, H. J. (2003). Protein Expression and Purification, 28, 331–339.CrossRefGoogle Scholar
  11. 11.
    Cha, H. J., Shin, H. S., Lim, H. J., Cho, H. S., Dalal, N. N., Pham, M. Q., et al. (2005). Biochemical Engineering Journal, 24, 225–233.CrossRefGoogle Scholar
  12. 12.
    Lim, H. J., & Cha, H. J. (2006). Enzyme and Microbial Technology, 39, 208–214.CrossRefGoogle Scholar
  13. 13.
    Yokomizo, A. Y., Jorge, S. A. C., Astray, R. M., Fernandes, I., Ribeiro, O. G., Horton, D. S. P. Q., et al. (2007). Journal of Biotechnology, 2, 102–109.CrossRefGoogle Scholar
  14. 14.
    Nilsen, S. A., & Castellino, F. J. (1999). Protein Expression and Purification, 16, 136–143.CrossRefGoogle Scholar
  15. 15.
    Park, J. H., Kim, H. Y., Han, K. H., & Chung, I. S. (1999). Enzyme and Microbial Technology, 25, 558–563.CrossRefGoogle Scholar
  16. 16.
    Deml, L., Wolf, H., & Wagner, R. (1999). Journal of Virological Methods, 79, 191–203.CrossRefGoogle Scholar
  17. 17.
    Jorge, S. A. C., Santos, A. S., Spina, A., & Pereira, C. A. (2008). Cytotechnology, 57, 51–59.CrossRefGoogle Scholar
  18. 18.
    Bovo, R., Galesi, A. L. L., Jorge, S. A. C., Piccoli, R. A. M., Moraes, A. M., Pereira, C. A., et al. (2008). Cytotechnology, 57, 23–35.CrossRefGoogle Scholar
  19. 19.
    Galesi, A. L. L., Pereira, C. A., & Moraes, A. M. (2007). Journal of Biotechnology, 2, 1399–1407.CrossRefGoogle Scholar
  20. 20.
    Batista, F. R. X., Pereira, C. A., Mendonça, R. Z., & Moraes, A. M. (2008). Cytotechnology, 57, 11–22.CrossRefGoogle Scholar
  21. 21.
    Galesi, A. L. L., Aguiar, M. A., Astray, R. M., Augusto, A. F. P., & Moraes, A. M. (2008). Cytotechnology, 57, 73–81.CrossRefGoogle Scholar
  22. 22.
    Swiech, K., Silva, C. S., Arantes, M. K., Yokomizo, A. Y., Astray, R. M., Pereira, C. A., et al. (2008). Biotechnology and Applied Biochemistry, 49, 41–49.CrossRefGoogle Scholar
  23. 23.
    Swiech, K., Rossi, N., Silva, B. G., Jorge, S. A. C., Astray, R. M., & Suazo, C. A. T. (2008). Cytotechnology, 57, 61–66.CrossRefGoogle Scholar
  24. 24.
    Batista, F. R. X., Moraes, A. M., Büntemeyer, H., & Noll, T. (2009). Biologicals, 37, 108–118.CrossRefGoogle Scholar
  25. 25.
    Hansen, B. E., Andersson, E. C., Madsen, L. S., Engberg, J., Sondergaard, L., Svejgaard, A., et al. (1998). Tissue Antigens, 51, 119–128.CrossRefGoogle Scholar
  26. 26.
    Hink, W. F. (1991). In Vitro Cellular & Developmental Biology, Animal, 27, 397–401.CrossRefGoogle Scholar
  27. 27.
    Schlaeger, E. J., Foggetta, M., Vonach, J. M., & Christensen, K. (1993). Biotechnology Techniques, 7, 183–188.Google Scholar
  28. 28.
    Batista, F. R. X., Pereira, C. A., Mendonça, R. Z., & Moraes, A. M. (2005). Cytotechnology, 49, 1–9.CrossRefGoogle Scholar
  29. 29.
    Batista, F. R. X., Pereira, C. A., Mendonça, R. Z., & Moraes, A. M. (2006). Electronic Journal of Biotechnology, 9, 522–532.CrossRefGoogle Scholar
  30. 30.
    Castle, P., & Robertson, J. S. (1999). Developments in Biological Standardization, 99, 191–196.Google Scholar
  31. 31.
    Zhao, L. S., Qin, S., Zhou, T. Y., Tang, H., Liu, L., & Lei, B. J. (2000). World Journal of Gastroenterology, 6, 239–243.Google Scholar
  32. 32.
    Freshney, R. I. (1994). Culture of animal cells: a manual of basic technique (3rd ed.). New York: Wiley.Google Scholar
  33. 33.
    Perrin, P., Lafon, M., & Sureau, P. (1996). In F. X. Meslin, M. M. Kaplan, & H. Koprowaski (Eds.), Laboratory techniques in rabies (pp. 383–388). Geneva: WHO.Google Scholar
  34. 34.
    Astray, R. M., Augusto, E., Yokomizo, A. Y., & Pereira, C. A. (2008). Journal of Biotechnology, 3, 98–103.CrossRefGoogle Scholar
  35. 35.
    Mendonça, R. Z., Palomares, L. A., & Ramírez, O. T. (1999). Journal of Biotechnology, 72, 61–75.CrossRefGoogle Scholar
  36. 36.
    Wang DI, Cooney CL, Demain AL, Dunnill P, Humphrey AE, Lilly MD (1979) in Fermentation and enzyme technology. John Wiley and Sons, New York, pp. 374Google Scholar
  37. 37.
    Echalier G (1997) in Drosophila Cells in culture. Academic Press, New York, pp. 1–67Google Scholar
  38. 38.
    Holzer, J. W., Mayrhofer, J., Leitner, J., Blum, M., Webersinke, G., Heuritsch, S., et al. (2003). Protein Expression and Purification, 29, 58–69.CrossRefGoogle Scholar
  39. 39.
    Hardy, E., Martínez, E., Diago, D., Díaz, R., González, D., & Herrera, L. (2000). Journal of Biotechnology, 77, 157–167.CrossRefGoogle Scholar
  40. 40.
    Sunil Kumar, G. B., Ganapathi, T. R., Revathi, C. J., Prasad, K. S. N., & Bapat, V. A. (2003). Protein Expression and Purification, 32, 10–17.CrossRefGoogle Scholar
  41. 41.
    Carvalhal, A. C., Coroadinha, A. S., Alves, P. A., Moreira, J. L., Hauser, H., & Carrondo, M. J. T. (2002). Enzyme and Microbial Technology, 30, 95–109.CrossRefGoogle Scholar
  42. 42.
    Mitsuhashi, J. (1989). In J. Mitsuhashi (Ed.), Invertebrate cell system applications (pp. 3–20). Tokio: CRC Press.Google Scholar
  43. 43.
    Rhiel, M. A., & Murhammer, D. W. (1995). Biotechnology and Bioengineering, 47, 640–650.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Fabiana R. X. Batista
    • 1
  • Kátia N. Greco
    • 2
  • Renato M. Astray
    • 2
  • Soraia A. C. Jorge
    • 2
  • Elisabeth F. P. Augusto
    • 3
  • Carlos A. Pereira
    • 2
  • Ronaldo Z. Mendonça
    • 2
  • Ângela M. Moraes
    • 1
    Email author
  1. 1.Departamento de Processos Biotecnológicos, Faculdade de Engenharia QuímicaUniversidade Estadual de CampinasCampinasBrazil
  2. 2.Laboratório de Imunologia ViralInstituto ButantanSão Paulo-SPBrazil
  3. 3.Laboratório de Biotecnologia IndustrialInstituto de Pesquisas Tecnológicas do Estado de São PauloSão Paulo-SPBrazil

Personalised recommendations