Nogueira Diogo E S, Rodrigues Carlos A V, Hashimura Yas, Jung Sunghoon, Lee Brian, Cabral Joaquim M S
Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
PBS Biotech, Camarillo, CA, USA.
Methods Mol Biol. 2021;2286:167-178. doi: 10.1007/7651_2020_287.
Human induced pluripotent stem cells (hiPSCs) have the potential to be used in a variety of biomedical applications, including drug discovery and Regenerative Medicine. The success of these approaches is, however, limited by the difficulty of generating the large quantities of cells required in a reproducible and controlled system. Bioreactors, widely used for industrial manufacture of biological products, constitute a viable strategy for large-scale production of stem cell derivatives. In this chapter, we describe the expansion of hiPSCs using the Vertical-Wheel™ bioreactor, a novel bioreactor configuration specifically designed for the culture of shear-sensitive cells. We provide protocols for the expansion of hiPSCs in suspension, both as floating aggregates and using microcarriers for cell adhesion. These methods may be important for the establishment of a scalable culture of hiPSCs, allowing the manufacturing of industrial- or clinical-scale cell numbers.
人诱导多能干细胞(hiPSCs)有潜力用于多种生物医学应用,包括药物发现和再生医学。然而,这些方法的成功受到在可重复和可控系统中生成所需大量细胞的困难的限制。广泛用于生物制品工业生产的生物反应器,是大规模生产干细胞衍生物的可行策略。在本章中,我们描述了使用Vertical-Wheel™生物反应器扩增hiPSCs的方法,该生物反应器是一种专门为培养对剪切力敏感的细胞而设计的新型生物反应器配置。我们提供了在悬浮状态下扩增hiPSCs的方案,既可以作为漂浮聚集体,也可以使用微载体促进细胞黏附。这些方法对于建立可扩展的hiPSCs培养体系可能很重要,从而能够生产工业规模或临床规模的细胞数量。