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Culturing Drosophila melanogaster (S2) in a chemostat

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Abstract

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.

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Abbreviations

CV:

Coefficient of variation

D:

Dilution rate

GLC:

Glucose concentration

GLN:

Glutamine concentration

mS :

Maintenance coefficient

µMax :

Maximum specific growth rate

NH4 :

Ammonium concentration

RVGP:

Rabies virus glycoprotein

S2:

Drosophila melanogaster cells lineage

X:

Viable cell concentration

YNH4/GLN :

Ammonium yield on glutamine

YNH4/X :

Ammonium yield on cells

YX/GLC :

Cell yield on glucose

YX/GLN :

Cell yield on glutamine

YX/S :

Global cell yield on substrate

Y trueX/S :

True cell yield on substrate

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Acknowledgments

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

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Authors and Affiliations

  1. Laboratório de Células Animais, Escola Politécnica––Universidade de São Paulo, P.O. Box 61.548, São Paulo, SP, CEP 05524-970, Brazil

    Paula Bruzadelle Vieira, Bruno Labate Vale da Costa & Aldo Tonso

  2. Universidade Federal de São Paulo––Campus São José dos Campos, Rua Talim, 330, São José dos Campos, SP, CEP 12231-280, Brazil

    Elisabeth de Fatima Pires Augusto

Authors
  1. Paula Bruzadelle Vieira
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  2. Bruno Labate Vale da Costa
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  3. Elisabeth de Fatima Pires Augusto
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  4. Aldo Tonso
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Correspondence to Aldo Tonso.

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Vieira, P.B., da Costa, B.L.V., de Fatima Pires Augusto, E. et al. Culturing Drosophila melanogaster (S2) in a chemostat. Biotechnol Lett 37, 533–538 (2015). https://doi.org/10.1007/s10529-014-1717-9

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  • DOI: https://doi.org/10.1007/s10529-014-1717-9

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