ALTEX :: Alternatives to Animal Experiments

2005, VOLUME 3

Evaluation of serum and animal protein-free media for the production of infectious bronchitis virus (M41) strain in a continuous cell line

Tereza C. Cardoso1, Maria C. B. Teixeira2, Natali Fachin2, Tatiani L. Camargo3, Deriane E. Gomes3, Anivaldo O. Corte Junior4 and Beatriz Anselmo5

1UNESP-Laboratório de Virologia, Curso de Medicina Veterinária, Faculdade de Odontologia, Universidade Estadual Paulista-Araçatuba, Br-São Paulo; 2Curso de Mestrado em Ciência Animal, UNESP-Campus de Araçatuba, Br-São Paulo; 3Fellowship Fundação de Amparo a Pesquisa do Estado de São Paulo-FAPESP, Br-São Paulo; 4Fellowship Conselho Nacional de Desenvolvimento Científico e Tecnológico-CNPq, Br-São Paulo; 5UNESP-Curso de Medicina Veterinária, Faculdade de Odontologia, Universidade Estadual Paulista - Araçatuba, Br-São Paulo

SUMMARY

Infectious bronchitis virus (IBV) is produced in vitro using specific pathogen free chicken embryos, primary chicken kidney cells (CKC) and/or tracheal organ culture (TOC). Regulatory authorities in Europe (EMEA) and in the United States (FDA) have encouraged biological manufactures to reduce or eliminate the use of live animals and/or products of animal origin in biological manufacturing processes. In this paper, a stable chicken embryo-related (CER) cell line was adapted and maintained in serum free (SF) or animal protein free (APF) medium after a direct switch of the medium. The most suitable media were Ex Cell 520 and Ex Cell 302. CER monolayers adapted to SF or APF were infected with IBV (M41 strain) in agitated suspended culture and the IBV titre obtained 48 h post infection was 2.8 x 104 PFU/ml in both cases. Thus, propagation of CER cells and culture of IBV can be performed without the use of animal serum or animal protein.

Keywords: IBV, serum free media, animal protein free media, virus production