Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production.
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 1x10(6) 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 1x10(6) cells/ml, at 3x10(6) 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.
Ferreira
Tiago B
TB
Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Apartado 12, P-2781-901, Oeiras, Portugal.
Ferreira
Ana L
AL
Carrondo
Manuel J T
MJ
Alves
Paula M
PM
D000256:Adenoviridae / Q000254:growth & development* / Q000302:isolation & purification*; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D016895:Culture Media, Serum-Free; D007668:Kidney / Q000196:embryology / Q000254:growth & development* / Q000821:virology*; D009994:Osmolar Concentration; D014776:Virus Cultivation / Q000379:methods*
D016895:Culture Media, Serum-Free