Bio-Safety Technology in Production of Bovine Herpesvirus Type 5 (BoHV-5) Using an Alternative Serum-Free Medium
Tereza C. Cardoso, Heitor F. Ferrari, Maria C.R. Luvizotto and Clarice W. Arns
DOI : 10.3844/ajbbsp.2007.125.130
American Journal of Biochemistry and Biotechnology
Volume 3, Issue 3
The detection and growth of bovine herpes virus type 5 (BoHV-5) was evaluated in three different media without foetal bovine serum or animal protein by infecting three different cell lines. The OPTI-PRO, VP-SFM and RPMI 1640 media were supplemented by L-glutamine, antibiotics added by 5Î¼g of insulin-like growth factor (IGF-I) and used to adapt CER, MDBK and CRIB cells in a statistic continuous culture system. The results obtained by CRIB and MDBK cells adaptation showed steady growth after 10th passages in OPTI-PRO medium, 20th passages in VP-SFM medium and 30th passages in RPMI 1640 medium, respectively. The OPTI-PRO and RPMI 1640 media were not able to support CER growth, even supplemented by IGF-1 and present apoptotic cells at 72h post-infection. The CER cells seeded after adaptation by 10th passages in VP-SFM medium added by 5Î¼g/20ml of IGF-1 growth factor revealed the best virus titres compared to MDBK and CRIB cell lines. It was concluded that CER, MDBK and CRIB cells infected by BoHV-5 serotype could be used for laboratory diagnosis propagated on much safer culture system of high biotechnology advances.
© 2007 Tereza C. Cardoso, Heitor F. Ferrari, Maria C.R. Luvizotto and Clarice W. Arns. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.