Comparative Analysis of MEM, RPMI 1640 Media on Rabies Virus Propagation in Vero Cells and Virus Quantification by FAT

Abstract

Rabies disease can be preventable through vaccination is the only way for effective control and the vaccine is administered as post exposure prophylaxis method (PEP) along with rabies immunoglobulin. For the preparation of the vaccine, the high titre rabies virus has to be propagated in suitable host system like Vero cells. In this study, the virus was propagated in tissue culture flask in two different tissue culture media namely MEM and RPMI-1640, keeping Vero cell line as the host system. This preliminary study is to find the media that yields a better viral titre as high, the influence of harvesting intervals on viral titre for the reason that the vaccine yield is directly proportional to the viral titre during the virus propagation stage and leads to cost effective vaccine. The Vero cell was revived from passage 154 and the part of cells were subjected for adaptation in RPMI-1640 media with gradual media replacement. The passaging of RPMI 1640 media cells were continued until it reaches the equal cell count of MEM (p-158). The confluent monolayer of Vero cells was maintained in the same passage level with appropriate media. The cell count of MEM media, RPMI-1640 were 10.28 x 106 and 10.35 x 106 respectively in 25cm2 tissue culture flasks. The Vero cells were sub-cultured in 175 cm2 tissue culture flasks infected with 0.2 MOI (Multiplicity of Infection) of virus and the viral titration was estimated through FAT test (Fluorescent Antibody Test). The highest viral titre obtained from RPMI-1640 media batch (10-6.125/ml) and MEM media batch (10-6.25 /ml). The two days interval viral harvests shows (Batch 1 & 3) the viral titre log ranged between 10-4.375 to 10-5.875 and three days interval harvests (Batch 2 & 4) the viral titre log ranged between 10-3.750 to 10-6.250 per mL.