School of Chemical and Bioprocess Engineering, and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
J Biotechnol. 2012 Oct 31;161(3):308-19. doi: 10.1016/j.jbiotec.2012.06.003. Epub 2012 Jun 30.
The continuous production of large numbers of red blood cells (RBCs) ex vivo is a challenging task due to process economics and complex culture conditions. In any serial passaging process, the culture conditions and operation mode are important criteria for achieving high proliferation with optimal passage lengths. The optimal inoculation concentration for serial passaging is a factor that affects both the kinetics and the total expansion performance. As part of our attempt to develop a scalable, economical and reproducible system for production of RBCs we used mathematical expressions to define the growth curves of peripheral blood derived erythroid progenitors over the course of their expansion process. We used a Gompertz function to evaluate the specific growth rate for the optimisation of inoculation concentration and passage lengths to achieve optimal expansion. This led to values of 3×10(5) cells/ml as the optimum inoculation concentration and 36 h as the optimum passage length. Also the variations in growth curves confirmed the altered growth kinetics of erythroid progenitors during sequential passaging in expansion process. Cost analysis suggested a 60-h passage length at every passage, resulting in a 42.9% process-cost reduction. However, this has increased the process duration in achieving the similar expansion factor. This methodology for optimising the expansion process of peripheral derived erythroid progenitors based on optimum culture conditions could provide us with a direction and an improved performance for scale-up applications.
由于工艺经济性和复杂的培养条件，体外大量连续生产红细胞（RBC）是一项具有挑战性的任务。在任何连续传代过程中，培养条件和操作模式都是实现高增殖和最佳传代数的重要标准。连续传代的最佳接种浓度是影响动力学和总扩增性能的因素之一。作为我们开发用于生产 RBC 的可扩展、经济和可重复的系统的一部分，我们使用数学表达式来定义外周血衍生的红系祖细胞在其扩增过程中的生长曲线。我们使用 Gompertz 函数来评估最佳接种浓度和传代数的特定生长率，以实现最佳扩增。这导致 3×10(5)个细胞/ml 作为最佳接种浓度，36 小时作为最佳传代时间。生长曲线的变化也证实了在扩增过程中连续传代时红系祖细胞的生长动力学发生了改变。成本分析表明，每传代 60 小时，可使工艺成本降低 42.9%。然而，这增加了实现类似扩增因子的过程持续时间。这种基于最佳培养条件优化外周血衍生红系祖细胞扩增过程的方法，可以为我们提供一个方向，并为扩大应用提供更好的性能。
