Journal of Biotechnology, 31 (1993) 1-15 1 © 1993 Elsevier Science Publishers B.V. All rights reserved 0168-1656/93/$06.00 BIOTEC 00944 Anticarsia gemmatalis nuclear polyhedrosis virus replication in serum-free and serum-reduced insect cell cultures Juan D. Claus, Gabriel E. Remondetto, Sergio A. Guerrero, Ana M. Demonte, Marcelo Murgula and Alberto J. Marcipar INTEBIO, Facultad de Bioqulmica y Ciencias Biol6gicas, Universidad Nacional del Litoral, Santa Fe, Argentina (Received 20 July 1992; revision accepted 29 September 1992) Summary In order to develop a financially feasible process to produce Anticarsia gem- matalis Nuclear Polyhedrosis virus in cell culture, we developed a lipidic supple- ment to replace fetal calf serum in insect cell culture media. The supplement, prepared with an extract of lipids from hen egg yolk, allowed us to reduce the contents of serum in the culture medium from 10% to 1%. IPLB-Sf-21 cells could be kept along consecutive passages in serum-reduced medium. The replication of AgNPV in HEYLE-supplemented cultures was evaluated. Extracellular virions production was the same as in FCS-supplemented-cultures, but the production level of polyhedral inclusion bodies was significantly lowered in HEYLE-supple- mented cultures. The reduced production of PIBs is related to a premature releasing of non-occluded particles as well as to a reduced synthesis of polyhedrin protein. Correspondence to: J.D. Claus, INTEBIO, Fac. Bioqulmica y Ciencias Biol6gicas,Universidad Nacional del Litoral, C.C. 530, Santa Fe, Repfiblica Argentina. Abbreviations: AcNPV, Autographa californica Nuclear Polyhedrosis virus; AgNPV, Anticarsia gem- matalis Nuclear Polyhedrosis virus; CLSE, commercial lipid supplement emulsion; DT, doubling time; FCS, fetal calf serum; HEYLE, hen egg yolk lipids emulsion; IPLB-Sf-21, Spodoptera frugiperda insect cells; MCD, maximum cell density; MOI, multiplicity of infection; NOV, non-occluded virus; NPV, Nuclear Polyhedrosis virus; pi, post-infection; PIB, polyhedral inclusion body; TCIDs0 , tissue culture infectious dose 50%.