Optimization of culture conditions for vaccinia virus propagation

Abstract

Background and Aim. Monkeypox virus has recently re emerged as one of the most significant zoonotic pathogens and a major global public health concern. Due to its genetic and antigenic similarity to other members of the Orthopoxvirus genus, the use of model viruses is essential for the development of vaccines and diagnostic approaches. In this context, Vaccinia virus is widely regarded as a key model system for Orthopoxvirus research and is extensively used in the development of vaccines and diagnostic tools. The aim of this study was to determine the optimal conditions for the cultivation of vaccinia virus.
Materials and Methods. Vaccinia virus was cultivated in Vero, HeLa, and LK cell lines. Viral replication activity was evaluated based on cytopathic effect observed in cell cultures and the level of viral biological activity. In addition, the effects of fetal bovine serum concentration, multiplicity of infection, and incubation temperature on viral replication were investigated. For each parameter, the efficiency of viral propagation was comparatively analyzed to determine optimal cultivation conditions.
Results. The results demonstrated that the highest level of viral replication was observed in HeLa cells. The optimal infection dose for efficient viral propagation was determined to be 0.00369 TCID₅₀/cell. Furthermore, 2% fetal bovine serum was sufficient to ensure stable and efficient viral growth. The most favorable incubation temperature was 37 °C, which supported the maintenance of viral biological activity.
Conclusion. The optimized cultivation conditions identified in this study enable efficient biomass production of vaccinia virus. These findings provide an important scientific basis for further fundamental and applied research on Orthopoxviruses, including the development of vaccines and diagnostic systems.