Production of small ruminant morbillivirus, rift valley fever virus and lumpy skin disease virus in CelCradle™ -500A bioreactors

Abstract

Background 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. 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). Results Two 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). Conclusions This 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.

Enhancing Dengue Virus Production and Immunogenicity with Celcradle™ Bioreactor: A Comparative Study with Traditional Cell Culture Methods

“…The Celcradle TM bioreactor system has been instrumental in supporting the production of various viruses, including Japanese encephalitis virus, hepatitis D virus-like particles, influenza A virus, small ruminant morbillivirus, rift valley fever virus, lumpy skin disease virus, and the current pandemic SARS-CoV-2 [ 11 , 13 , 32 , 33 , 34 , 35 ]. In this study, we successfully produced DENV using suspension-adapted Vero cells in the CelCradle TM bioreactor system.…”

High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor

“…The CelCradle™ bioreactor (Esco Aster Pte. Ltd., Singapore, Singapore) was previously reported to support efficient production of Japanese encephalitis virus [33], hepatitis D virus-like particles [34], different strains of influenza A virus [35], severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) [36], as well as a number of viruses relevant for veterinary vaccine purposes [37]. It is a packed-bed bioreactor using BioNOCII™ macrocarriers and it employs a tide-motion principle, thereby conferring minimal shear stress to cells during medium circulation.…”