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Growth, metabolism and baculovirus production in suspension cultures of an Anticarsia gemmatalis cell line

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Abstract

The UFL-AG-286 cell line, established from embryonic tissue of the lepidopteran insect Anticarsia gemmatalis, has been identified as a good candidate to be used as a cellular substrate in the development of a process for in vitro production of the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus, a baculovirus widely used as bioinsecticide. In order to characterize the technological properties of this cell line and evaluate its feasibility to use it for the large-scale production of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus, UFL-AG-286 cells were adapted to grow as agitated suspension cultures in spinner-flasks. Batch suspension cultures of adapted cells in serum-supplemented TC-100 medium grew with a doubling time of about 29 h and reached a maximum cell density higher than 3.5 × 106 viable cells ml−1. At the end of the growth period glucose was completely depleted from the culture medium, but l-lactate was not produced. Amino acids, with the exception of glutamine, were only negligibly consumed or produced. In contrast to other insect cell lines, UFL-AG-286 cells appeared to be unable to synthesize alanine as a metabolic way to dispose the by-product ammonia. The synchronous infection of suspension cultures with Anticarsia gemmatalis multicapsid nucleopolyhedrovirus in the early to medium exponential growth phase yielded high amounts of both viral progenies per cell and reduced the specific demands of UFL-AG-286 cells for the main nutrients.

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Abbreviations

AgMNPV:

Anticarsia gemmatalis multicapsid nucleopolyhedrovirus

NOV:

non-occluded virus

PIB:

polyhedral inclusion body

TCID50% :

tissue culture infectious dose 50%

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Acknowledgements

The authors gratefully acknowledge support from ANPCyT (PICT 08-04411 BID 802 OC/AR) and Universidad Nacional del Litoral (CAI + D 2002).

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Authors and Affiliations

  1. Laboratorio de Virología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, CC 242, S3000ZAA, Santa Fe, República Argentina

    Verónica Viviana Gioria & Juan Daniel Claus

  2. Helmholtz Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124, Braunschweig, Germany

    Volker Jäger

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  1. Verónica Viviana Gioria
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  2. Volker Jäger
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  3. Juan Daniel Claus
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Correspondence to Juan Daniel Claus.

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Gioria, V.V., Jäger, V. & Claus, J.D. Growth, metabolism and baculovirus production in suspension cultures of an Anticarsia gemmatalis cell line. Cytotechnology 52, 113–124 (2006). https://doi.org/10.1007/s10616-006-9042-3

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