title: Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production.
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 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.
authors.affiliation: Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Apartado 12, P-2781-901, Oeiras, Portugal.
authors.forename: Tiago B
authors.initials: TB
authors.lastname: Ferreira
authors.forename: Ana L
authors.initials: AL
authors.lastname: Ferreira
authors.forename: Manuel J T
authors.initials: MJ
authors.lastname: Carrondo
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
mesh_terms: 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*
chemical_list: D016895:Culture Media, Serum-Free