Introduction: Rotavirus disease is associated with 600.000 deaths worldwide among children younger than five years age. Most symptomatic episodes occur in young children between the ages of 3 months and 2 years. The development of rotavirus vaccine is very important to prevent the severe rotavirus disease. This vaccine is prepared with rotavirus suspensions obtained of cell cultures infected with different rotavirus serotypes.
Objectives: The objective of this study was to evaluate the potency rotavirus suspension produced in bioreactor (large scale) the serotypes G1, G2, G3, G4 and G9.
Methods and Materials: Vero cells added on solid microcarriers (cytodex 1) were cultivated with serum free- medium in bioreactor of 5L (Celligem NBS) with initial concentration cellular of 1, 0 x 109 cells. After three or four days (100% cell confluence) the cells were infected with different rotavirus serotypes (reassortant strains G1, G2, G3, G4 and G9 from NIH). Samples this supernatant was taken for observed cytopatic effects and cellular concentration. When observed 90-100% of cytopatic effects, the harvests of the supernatant of these cultures were realized and the samples were taken to determine the rotavirus titers by PFU (Plate Forming Units) test. For this test were utilized MA-104 cells cultivated in plate six wells.
Results: The titers obtained in the samples of the harvest were 106.2, 106.3, 106.2, 106.0 and
107, 5 PFU/ml for G1, G2, G3, G4 and G9 rotavirus serotypes respectively.
Discussion: The titers found in the supernatant of the cultures infected with different rotavirus serotypes showed that the utilization of Vero cell added on solid microcarriers cultivated in bioreactor is a good strategy to optimize the rotavirus production at Institute Butantan.
Financial support: Butantan Foundation