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Stability of Minimum Essential Medium functionality despite l-glutamine decomposition

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Abstract

l-Glutamine (l-Gln) instability in liquid media is a well-known fact. Also, negative effect of ammonia, one of the l-Gln degradation products, on viability of many cell cultures and on replication of different viruses has been described. However, negative effects of ammonia have been reported in doses excessively exceeding those that could be generated in regularly used liquid culture media due to spontaneous l-Gln breakdown (below 2 mM). Traditional virus vaccine production processes have been established and registered involving l-Gln containing media use. Eventual culture media replacement in the regular production process belongs to the major regulative changes that require substantial financial expenses. The aim of this study was to evaluate the effect of storage of Minimum Essential Media with Hanks salts on their relevant biological functions during virus vaccine production process in relation to l-Gln decrease. Our results show a cell type dependent effect of spontaneous l-Gln degradation during medium storage. They also suggest that for cell cultures used in measles, mumps, and rubella virus production the media retain their functionality in respect to cell viability or virus growth over a certain time window despite l-Gln degradation.

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Acknowledgments

The authors wish to thank Mrs. Tamara Božić, MSc, and the Virus Vaccine Quality Control Unit as well as Mrs. Vesna Feletar, MS, and Mrs. Marija Živković, MS, from the Virus Vaccine Production Preparation Unit of the Institute of Immunology Inc., for their participation and collaboration in certain parts of this study. Also, we are grateful to Mr. Nediljko Pavlović, MSc, for his help in data analysis. The work was financed by the Institute of Immunology, Inc.

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Authors and Affiliations

  1. Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, 10 000, Zagreb, Croatia

    Maja Jagušić, Dubravko Forčić, Marija Brgles, Maja Šantak, Mladen Jergović, Krešo Bendelja & Beata Halassy

  2. Institute of Immunology Inc., Rockefellerova 10, 10 000, Zagreb, Croatia

    Leonida Kutle & Ljerka Kotarski

  3. Center of Excellence for Virus Immunology and Vaccines, CERVirVac, Zagreb, Croatia

    Maja Jagušić, Dubravko Forčić, Marija Brgles, Maja Šantak, Mladen Jergović, Krešo Bendelja & Beata Halassy

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  1. Maja Jagušić
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  2. Dubravko Forčić
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  3. Marija Brgles
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  4. Leonida Kutle
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  6. Mladen Jergović
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  8. Krešo Bendelja
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  9. Beata Halassy
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Correspondence to Beata Halassy.

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Jagušić, M., Forčić, D., Brgles, M. et al. Stability of Minimum Essential Medium functionality despite l-glutamine decomposition. Cytotechnology 68, 1171–1183 (2016). https://doi.org/10.1007/s10616-015-9875-8

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  • DOI: https://doi.org/10.1007/s10616-015-9875-8

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