Comparative analysis of reovirus replication efficiency in HEK293T, L929, and huh-7 cell lines: implications for reovirus propagation in oncolytic therapy.

Abstract

Oncolytic viruses, such as reovirus, represent a transformative approach in cancer therapy by selectively targeting malignant cells through mechanisms like RAS pathway activation. While clinical trials have demonstrated the safety and efficacy of reovirus in combination therapies, efficient large-scale production remains a critical challenge. Optimal cell lines for maximizing reovirus replication are essential for scalable manufacturing but remain understudied. This study evaluates reovirus propagation in HEK293T, L929, and Huh-7 cell lines to identify the most effective platform for large-scale production. Reovirus replication and cytotoxicity were assessed using plaque assays, trypan blue viability staining, real-time PCR, and MTT assays. Cell lines were infected at a MOI of 10, with samples collected at 0, 24, 48, and 72 h post-infection. HEK293T cells showed the strongest cytopathic effects after reovirus infection compared to L929 and Huh-7 cells. Trypan blue staining confirmed reduced viability in HEK293T cells. Plaque assays indicated higher viral replication in these cells. Real-time PCR revealed increased viral RNA levels in HEK293T samples. MTT assays demonstrated decreased metabolic activity, reflecting greater virus-induced toxicity. HEK293T cells emerge as the optimal platform for reovirus production due to their robust replication capacity and pronounced cytotoxicity, key attributes for oncolytic therapy applications. This study underscores HEK293T's potential to meet the increasing demand for scalable oncolytic virus manufacturing, paving the way for enhanced therapeutic applications in cancer treatment.