Biotechnology Letters

, Volume 37, Issue 3, pp 533–538 | Cite as

Culturing Drosophila melanogaster (S2) in a chemostat

  • Paula Bruzadelle Vieira
  • Bruno Labate Vale da Costa
  • Elisabeth de Fatima Pires Augusto
  • Aldo Tonso
Original Research Paper


Insect cells are used for the expression of complex proteins in products such as vaccines and biopharmaceuticals. Physiology of a Drosophila melanogaster (lineage S2), transfected to stably express rabies virus glycoprotein (RVGP), was studied in batch culture and in a chemostat with serum-free medium. In batch mode, the system reached 3 × 107 cells mL−1 with specific growth rate of 1.5 d−1 with RVGP at 2.50 µg L−1. When operated continuously, three well-defined steady states were achieved at dilution rates (D) of 0.8, 0.5 and 0.2 d−1. The residual glucose and glutamine concentrations and the cell yields on glucose and glutamine decreased at lower D. High values of substrate consumption for maintenance may explain this variation in yields. The results indicated that glucose is not the limiting substrate of this process.


Bioprocess engineering Bioreactor Continuous culture Drosophila melanogaster Insect cell Rabies virus glycoprotein 

List of abbreviations


Coefficient of variation


Dilution rate


Glucose concentration


Glutamine concentration


Maintenance coefficient


Maximum specific growth rate


Ammonium concentration


Rabies virus glycoprotein


Drosophila melanogaster cells lineage


Viable cell concentration


Ammonium yield on glutamine


Ammonium yield on cells


Cell yield on glucose


Cell yield on glutamine


Global cell yield on substrate


True cell yield on substrate



This work was supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico).


  1. Batista FRX, Moraes AM, Büntemeyer H, Noll T (2009) Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium. Biologicals 37:108–118CrossRefPubMedGoogle Scholar
  2. Bedard C, Tom R, Kamen A (1993) Growth nutrient consumption and end product accumulation in Sf9 and BTI-EAA insect cell cultures: insights into growth limitation and metabolism. Biotechnol Prog 9:615–624CrossRefPubMedGoogle Scholar
  3. Bovo R, Galesi ALL, Jorge SAC, Piccoli RAM et al (2008) Kinetic response of a Drosophila melanogaster cell line to different medium formulations and culture conditions. Cytotechnology 57:23–35CrossRefPubMedCentralPubMedGoogle Scholar
  4. Chisti Y (2001) Hydrodynamic damage to animal cells critical. Crit Rev Biotechnol 21:67–110CrossRefPubMedGoogle Scholar
  5. Compton ME (2011) Elements of in vitro research. In: Trigiano RN, Gray DJ (eds) Plant tissue culture, development, and biotechnology. CRC Press, Boca Raton, pp 57–64Google Scholar
  6. Galesi ALL, Aguiar MA, Astray RM, Augusto EFP et al (2008) Growth of recombinant Drosophila melanogaster Schneider 2 cells producing rabies virus glycoprotein in bioreactor employing serum-free medium. Cytotechnology 57:73–81CrossRefPubMedCentralPubMedGoogle Scholar
  7. Hoskisson PA, Hobbs G (2005) Continuous culture––making a comeback? Microbiology 151:3153–3159CrossRefPubMedGoogle Scholar
  8. Ikonomou L, Schneider YJ, Agathos SN (2003) Insect cell culture for industrial production of recombinant proteins. Appl Microbiol Biotechnol 62:1–20CrossRefPubMedGoogle Scholar
  9. Jorge SA, Santos AS, Spina A, Pereira CA (2008) Expression of the hepatitis B virus surface antigen in Drosophila S2 cells. Cytotechnology 57:51–59CrossRefPubMedCentralPubMedGoogle Scholar
  10. Lim HJ, Cha HJ (2006) Observation and modeling of induction effect on human transferring production from stably transfected Drosophila S2 cell culture. Enzym Microb Technol 39:208–214CrossRefGoogle Scholar
  11. Moraes AM, Jorge SAC, Astray RM, Suazo CAT et al (2012) Drosophila melanogaster S2 cells for expression of heterologous genes: from gene cloning to bioprocess development. Biotechnol Adv 30:613–628CrossRefPubMedGoogle Scholar
  12. Pamboukian MM, Jorge SAC, Santos MG, Yokomizo AY et al (2008) Insect cells respiratory activity in bioreactor. Cytotechnology 57:37–44CrossRefPubMedCentralPubMedGoogle Scholar
  13. Pirt SJ (1965) The maintenance energy of bacteria in growing. Proc R Soc Lond B Biol Sci 163:224–231CrossRefPubMedGoogle Scholar
  14. Swiech K, Rossi N, Astray RM, Suazo CAT (2008) Enhanced production of recombinant rabies virus glycoprotein (rRVGP) by Drosophila melanogaster S2 cells through control of culture conditions. Cytotechnology 57:67–72CrossRefPubMedCentralPubMedGoogle Scholar
  15. Ventini DC, Astray RM, Lemos MAN, Jorge SAC et al (2010) Recombinant rabies virus glycoprotein synthesis in bioreactor by transfected Drosophila melanogaster S2 cells carrying a constitutive or an inducible promoter. J Biotechnol 146:169–172CrossRefPubMedGoogle Scholar
  16. Yokomizo A, Jorge S, Astray R, Fernandes I et al (2007) Rabies virus glycoprotein expression in Drosophila S2 cells. I. Functional recombinant protein in stable co-transfected cell line. Biotechnol J 2:102–109CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Paula Bruzadelle Vieira
    • 1
  • Bruno Labate Vale da Costa
    • 1
  • Elisabeth de Fatima Pires Augusto
    • 2
  • Aldo Tonso
    • 1
  1. 1.Laboratório de Células AnimaisEscola Politécnica––Universidade de São PauloSão PauloBrazil
  2. 2.Universidade Federal de São Paulo––Campus São José dos CamposSão José dos CamposBrazil

Personalised recommendations