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Cytotechnology

, Volume 57, Issue 1, pp 23–35 | Cite as

Kinetic response of a Drosophila melanogaster cell line to different medium formulations and culture conditions

  • R. Bovo
  • A. L. L Galesi
  • S. A. C. Jorge
  • R. A. M. Piccoli
  • A. M. Moraes
  • C. A. Pereira
  • E. F. P. Augusto
SI:Heterologous Gene Expression

Abstract

In the past few years, Drosophila melanogaster cells have been employed for recombinant protein production purposes, and a comprehensive knowledge of their metabolism is essential for process optimization. In this work, the kinetic response of a Schneider S2 cell line, grown in shake flasks, in two different culture media, the serum-free SF900-II® and the serum-supplemented TC-100, was evaluated. Cell growth, amino acids and glucose uptake, and lactate synthesis were measured allowing the calculation of kinetic parameters. The results show that S2 cells metabolism was able to adjust to different environmental situations, as determined by medium formulation, as well as by the particular situation resulting from the culture conditions. Cells attained a 163% higher final cell concentration (1.4 × 107 cells mL−1) in SF900 II® medium, when compared to serum-supplemented TC-100 medium. Also, a maximum specific cell growth rate 52% higher in SF900 II® medium, when compared to serum-supplemented TC-100 one, was observed. Glutamine was the growth limiting factor in SF900 II® medium, while glucose, sometimes associated with glutamine, controlled growth in serum-supplemented TC-100 medium based formulation. The different pattern of lactate production is an example of the versatility of the metabolism of these cells. This by-product was produced only in glutamine limitation, but the amount synthesized depended not only on the excess glucose, but on other medium components. Therefore, in serum-supplemented TC-100 medium a much smaller lactate amount was generated. Besides, glucose was identified not only as a growth limiting factor, but also as a viability limiting factor, since its depletion accelerated cell death.

Keywords

S2 cells Metabolism Glucose Glutamine Lactate Alanine Kinetic parameters SF900 II® TC-100 

Notes

Acknowledgments

This work was supported in part by grants from FAPESP, CNPq, and Fundação Butantan. A.M. Moraes and C. A. Pereira are the recipients of a CNPq research fellowship.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • R. Bovo
    • 1
  • A. L. L Galesi
    • 2
  • S. A. C. Jorge
    • 1
  • R. A. M. Piccoli
    • 3
  • A. M. Moraes
    • 2
  • C. A. Pereira
    • 1
  • E. F. P. Augusto
    • 3
  1. 1.Laboratório de Imunologia ViralInstituto ButantanSão PauloBrazil
  2. 2.Departamento de Processos Biotecnológicos, Faculdade de Engenharia QuímicaUniversidade Estadual de CampinasCampinasBrazil
  3. 3.Laboratório de Biotecnologia IndustrialInstituto de Pesquisas Tecnológicas—IPTSão PauloBrazil

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