Laboratory of functional and environmental ecology, Faculty of Sciences and Technology Sidi Mohammed Ben Abdellah University, Imouzzer Road, BP, 2202, Fez, Morocco.
Laboratory of Research and Development virology, MCI Animal Health, Lot. 157, Zone Industrielle Sud-Ouest (ERAC) B.P: 278, 28810, Mohammedia, Morocco.
BMC Vet Res. 2021 Feb 27;17(1):93. doi: 10.1186/s12917-021-02801-4.
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).
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 × 10 using the CelCradle™-500A with a shorter doubling time as compared to 9.45 × 10 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 × 10 as compared to 6.3 × 10 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).
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.
动物疫苗接种是阻止对牲畜造成巨大损害的疾病传播的重要方法，也是经济损失和潜在向人类传播的重要方法。因此，使用简单廉价的生物加工解决方案生产有效疫苗的方法非常重要。目前使用的传统培养系统在劳动力和时间方面都不经济。此外，它们为细胞生长提供的表面积有限。在这项研究中，CelCradle™-500A 被评估为替代目前使用的传统培养系统（如细胞工厂）的一种选择，用于生产小反刍动物麻疹病毒 (PPR)、裂谷热病毒 (RVF) 和牛结节性皮肤病病毒 (LSD) 的病毒疫苗。
使用两种类型的细胞——Vero 细胞和原代羔羊睾丸细胞来生产这些病毒。这项研究分为两个阶段进行：a）细胞生长的优化和 b）病毒培养。与 Cell factories 相比，使用 CelCradle™-500A 可将 Vero 细胞培养到更高的细胞密度 3.04×10，倍增时间更短。与在 Cell factories 中接种相比，这代表细胞数量增加了 19 倍，而在 Cell factories 中仅增加了 3.7 倍。LT 细胞的细胞密度略高，为 6.7×10，而 Cell factories 中的细胞密度为 6.3×10。与 Cell factories 相比，这些细胞的密度变化倍数在 CelCradle™-500A 中为 3 倍，在 Cell factories 中为 2.5 倍。传统系统和生物反应器中的滴度没有显著差异。然而，裂谷热病毒和牛结节性皮肤病病毒的细胞特异性病毒产量更高（裂谷热病毒为 25 个病毒/细胞，牛结节性皮肤病病毒为 21.9 个病毒/细胞，而裂谷热病毒为 19.9 个病毒/细胞，牛结节性皮肤病病毒为 10 个病毒/细胞）。
这项工作代表了在 CellCradle™-500A 生物反应器中进行原代羔羊睾丸细胞培养的一项新研究。此外，由于获得了高细胞密度并且可以线性扩展，因此可以使用其他培养工艺（如灌注）进一步优化滴度。
