Division of Electrical and Computer Engineering, Louisiana State University, Baton Rouge, Louisiana.
Bioengineering Graduate Program, University of Notre Dame, Notre Dame, Indiana.
Biotechnol Bioeng. 2018 Jul;115(7):1717-1728. doi: 10.1002/bit.26601. Epub 2018 Apr 10.
With recent advances in biotechnology, mammalian cells are used in biopharmaceutical industries to produce valuable protein therapeutics and investigated as effective therapeutic agents to permanently degenerative diseases in cell based therapy. In these exciting and actively expanding fields, a reliable, efficient, and affordable platform to culture mammalian cells on a large scale is one of the most vital necessities. To produce and maintain a very large population of anchorage-dependent cells, a microcarrier-based stirred tank bioreactor is commonly used. In this approach, the cells are exposed to harmful hydrodynamic shear stress in the bioreactor and the mass transfer rates of nutrients and gases in the bioreactor are often kept below an optimal level to prevent cellular damages from the shear stress. In this paper, a hollow microcarrier (HMC) is presented as a novel solution to protect cells from shear stress in stirred bioreactors, while ensuring sufficient and uniform mass transfer rate of gases and nutrients. HMC is a hollow microsphere and cells are cultured on its inner surface to be protected, while openings on the HMC provide sufficient exchange of media inside the HMC. As a proof of concept, we demonstrated the expansion of fibroblasts, NIH/3T3 and the expansion and cardiac differentiation of human induced pluripotent stem cells, along with detailed numerical analysis. We believe that the developed HMC can be a practical solution to enable large-scale expansion of shear-sensitive anchorage-dependent cells in an industrial scale with stirred bioreactors.
随着生物技术的最新进展，哺乳动物细胞被用于生物制药行业生产有价值的蛋白质治疗药物，并被研究作为细胞治疗中永久性退行性疾病的有效治疗剂。在这些令人兴奋和积极扩展的领域中，一个可靠、高效且经济实惠的大规模培养哺乳动物细胞的平台是最关键的需求之一。为了生产和维持非常大量的锚定依赖性细胞，通常使用基于微载体的搅拌槽生物反应器。在这种方法中，细胞在生物反应器中暴露于有害的流体剪切力下，并且生物反应器中的营养物质和气体的传质速率通常保持在低于最佳水平以下，以防止细胞因剪切力而受损。在本文中，提出了一种中空微载体（HMC）作为一种保护细胞免受搅拌生物反应器中剪切力的新型解决方案，同时确保气体和营养物质的充分和均匀传质速率。HMC 是一种中空微球，细胞在其内部表面上进行培养以得到保护，而 HMC 上的开口提供了 HMC 内部介质的充分交换。作为概念验证，我们展示了成纤维细胞、NIH/3T3 的扩增以及人诱导多能干细胞的扩增和心脏分化，同时进行了详细的数值分析。我们相信，开发的 HMC 可以成为一种实用的解决方案，能够在搅拌生物反应器中实现大规模扩增对剪切力敏感的锚定依赖性细胞。
