Analysis of Continuous Cell Lines Vero, BHK21, and RK13 as Alternatives to MRC5 for Rubella Virus Propagation: A Study on Cell Proliferation and Titer Virus
Keywords:
continuous cell line, MRC5, BHK21, Vero, RK13, cell proliferation, virus titration, rubellaAbstract
Rubella virus remains a relevant global health concern, particularly due to its teratogenic effects when infection occurs during early pregnancy, potentially leading to congenital rubella syndrome. In vaccine development, the choice of host cell substrate plays a crucial role in ensuring both the safety and scalability of production. Currently, MRC5 human diploid fibroblast cells are widely utilized; however, they present certain limitations, including a finite lifespan and ethical concerns regarding their origin. This research aims to investigate the feasibility of three continuous cell lines, Vero, BHK21, and RK13, as potential alternatives to MRC5 for rubella virus cultivation. The study encompasses analysis of proliferation studies (morphology, viability, cell density, and population doubling time/PDT) and viral titer quantification post-infection. Each cell line was assessed for morphology, viability, cell density, and population doubling time (PDT). The most promising candidate based on proliferation characteristics was further tested for its ability to support rubella virus replication. Viral titers were measured using CCID₅₀ methods. The results of this study demonstrated that each cell line exhibited distinct morphological characteristics reflecting their tissue of origin, with excellent viability levels averaging above 95%. Among the candidates, Vero cells demonstrated the highest average cell density and the most favorable PDT value (1.6 days), warranting their inclusion in the subsequent virus inoculation phase. Rubella virus propagated in Vero cells achieved a titer of 5.53 logCCID₅₀/mL on day 14, which was higher than the 4.53 logCCID₅₀/mL observed in the MRC5 control group. Vero cells demonstrated strong potential as an alternative to MRC5 for rubella virus production, offering advantages in terms of proliferation, viral yield, and long-term sustainability. These findings contribute valuable insights for the development of more sustainable, scalable, and ethically sourced rubella vaccines, supporting global immunization initiatives.
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