Two Different Serum-free Media and Osmolality Effect Upon Human 293 Cell Growth and Adenovirus Production |
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Authors: | Tiago?B?Ferreira Ana?L?Ferreira Manuel?J?T?Carrondo Email author" target="_blank">Paula?M?AlvesEmail author |
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Institution: | (1) Instituto de Biologia Experimental e Tecnológica (IBET)/Instituto de Tecnologia Química e Biológica (ITQB), Apartado 12, P-2781-901 Oeiras, Portugal;(2) Laboratório de Engenharia Bioquímica, Faculdade de Ciências e Tecnologia (FCT), Universidade Nova de Lisboa (UNL), P-2825-114 Monte da Caparica, Portugal |
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Abstract: | Adenoviruses are promising vectors for gene therapy and vaccination protocols. Consequently, the market demands for adenovirus
are increasing, driving the search for new methodologies for large-scale production of concentrated vectors with warranted
purity and efficacy, in a cost-effective way. Nevertheless, the production of adenovirus is currently limited by the so-called
‘cell density effect’, i.e. a drop in cell specific productivity concomitant with increased cell concentration at infection.
Of two different serum-free culture media (CD293 and EX-Cell), evaluated for their effect on human 293 cells growth and adenovirus
production at cell densities higher than 1×106 cells/ml, EX-Cell proved the better medium for cell growth. Although adenovirus production was equivalent in both media when
the infection was performed at 1×106 cells/ml, at 3×106 cells/ml CD293 was the better. This result related to the high ammonia content in EX-Cell medium at the highest cell concentration
at infection. Besides this, the large-scale production of these vectors at high cell densities often requires re-feed strategies,
which increase medium osmolality. While a negative effect on cell growth was observed with increasing osmolalities, adenovirus
productivity was only affected for osmolalities higher than 430 mOsm.
Revisions requested 24 August 2005; Revisions received 9 September 2005 |
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Keywords: | adenovirus high cell densities human 293 cello osmolality serum-free media |
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