International Journal of Applied Science - Research and Review Open Access

  • ISSN: 2394-9988
  • Journal h-index: 10
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Abstract

Sequential Adaptation of Vero Cell Lines in Serum Free Medium for Fixed Rabies Virus Propagation

Jemal Mohammed1,2*, Abebe Mengesha1 , Birhanu Hurisa1 , Demise Mulugeta1 , Abate Waldetensai1 , Yeweynshet Tesera1 , Worku Gemechu1 , Ayele Bizuneh1 , Dejene Getachaw1 and Asnake Desalegn2

Background: Vero cells are derived from kidney of the African green monkey (Cercopithecus aethiops) and proliferated in artificial growth medium. They are used as substrate for rabies virus propagation to produce cell culture based vaccines; however serum supplemented medium to proliferate Vero cells are very expensive and source of different contaminant agents that affect the quality and affordability products.

Objectives: The aim of this research was to adapt Vero cell lines in serum free medium through sequential serum reduction for propagation of Pasteur virus (PV) and Evelyn Rokitnicki Abelseth (ERA) rabies viruses.

Methods: Vero cells were proliferated in various serum concentration supplemented medium by gradual reducing from 10% to 0% serum concentration. Viable cells were counted and sub- cultured to passage seven in each serum concentrations. PV and ERA rabies were used to infect the cells proliferated in each serum concentration based on their multiplicity of infection and Virus titers of both viruses were calculated and expressed as tissue culture infectivity dose fifty(TCID50).

Results: The maximum viable cells density of Vero cells at each serum supplemented medium (0%, 1%, 2.5%, 5%, 7.5% and 10%) were 2.85 × 106 cells/ml, 2.75 × 106 cells/ml, 2.75 × 106 cells/ml, 2.70 × 106 cells/ml, 2.92 × 106 cells/ml and 2.75 × 106 cells/ml, respectively. The maximum virus titers were 105.36 TCID50/ml and 105.61 TCID50/ml for PV and ERA respectively in 0% serum concentration medium proliferated cells.

Conclusion: From the results of this study; it can be concluded that Vero cells adapted in serum free conditions were produce sufficient rabies virus titers propagation used for anti-rabies vaccine production.