Abstract
BackgroundAnimal vaccination is an important way to stop the spread of diseases causing immense damage to livestock and economic losses and the potential transmission to humans. Therefore effective method for vaccine production using simple and inexpensive bioprocessing solutions is very essential. Conventional culture systems currently in use, tend to be uneconomic in terms of labor and time involved. Besides, they offer a limited surface area for growth of cells. In this study, the CelCradle™-500A was evaluated as an alternative to replace conventional culture systems in use such as Cell factories for the production of viral vaccines against small ruminant morbillivirus (PPR), rift valley fever virus (RVF) and lumpy skin disease virus (LSD).ResultsTwo types of cells Vero and primary Lamb Testis cells were used to produce these viruses. The study was done in 2 phases as a) optimization of cell growth and b) virus cultivation. Vero cells could be grown to significantly higher cell densities of 3.04 × 109 using the CelCradle™-500A with a shorter doubling time as compared to 9.45 × 108 cells in Cell factories. This represents a 19 fold increase in cell numbers as compared to seeding vs only 3.7 fold in Cell factories. LT cells achieved modestly higher cell densities of 6.7 × 108 as compared to 6.3 × 108 in Cell factories. The fold change in densities for these cells was 3 fold in the CelCradle™-500A vs 2.5 fold in Cell factories. The titers in the conventional system and the bioreactor were not significantly different. However, the Cell-specific virus yield for rift valley fever virus and lumpy skin disease virus are higher (25 virions/cell for rift valley fever virus, and 21.9 virions/cell for lumpy skin disease virus versus 19.9 virions/cell for rift valley fever virus and 10 virions/cell for lumpy skin disease virus).ConclusionsThis work represents a novel study for primary lamb testis cell culture in CellCradle™-500A bioreactors. In addition, on account of the high cell densities obtained and the linear scalability the titers could be further optimized using other culture process such us perfusion.
Highlights
Animal vaccination is an important way to stop the spread of diseases causing immense damage to livestock and economic losses and the potential transmission to humans
This work represents a novel study for primary lamb testis cell culture in CellCradleTM-500A bioreactors
We evaluated the use of the CelCradleTM -500A bioreactor for the cultivation of Small ruminant morbillivirus or peste des petits ruminants (PPR), Rift valley fever (RVF) and Lumpy skin disease of cattle (LSD) viruses using two types of cells; Vero cells for PPR cultivation and rift valley fever virus (RVF) viruses and primary lamb testis cells (LT) for lumpy skin disease virus (LSD) virus
Summary
Animal vaccination is an important way to stop the spread of diseases causing immense damage to livestock and economic losses and the potential transmission to humans. Conventional culture systems currently in use, tend to be uneconomic in terms of labor and time involved They offer a limited surface area for growth of cells. They are time consuming and involve heavy operations Microcarriers such as Cytodex have been largely used for adherent cells, offering good mixing and oxygen transfer but often resulting in low cell densities due to accumulation of toxic metabolites and a high shear stress. To alleviate the shear stress problem, other reactors have been developed such as hollow fiber [1], packed-bed bioreactors [2] and Wave bioreactors [3] They have the advantages of good mixing, aeration and nutrient supply, but require an exterior oxygenation system and sophisticated operation skills
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