Critical Analytics for Manufacturing of Personalized Medicine Interdisciplinary Research Group, Singapore-MIT Alliance in Research and Technology, Singapore, Singapore.
Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
Biotechnol Bioeng. 2021 May;118(5):1951-1961. doi: 10.1002/bit.27709. Epub 2021 Mar 1.
Medium perfusion is critical in maintaining high cell concentration in cultures. The conventional membrane filtration method for medium exchange has been challenged by the fouling and clogging of the membrane filters in long-term cultures. In this study, we present a miniature auto-perfusion system that can be operated inside a common-size laboratory incubator. The system is equipped with a spiral microfluidic chip for cell retention to replace conventional membrane filters, which fundamentally overcomes the clogging and fouling problem. We showed that the system supported continuous perfusion culture of Chinese hamster ovary (CHO) cells in suspension up to 14 days without cell retention chip replacement. Compared to daily manual medium change, 25% higher CHO cell concentration can be maintained at an average auto-perfusion rate of 196 ml/day in spinner flask at 70 ml working volume (2.8 VVD). The auto-perfusion system also resulted in better cell quality at high concentrations, in terms of higher viability, more uniform and regular morphology, and fewer aggregates. We also demonstrated the potential application of the system for culturing mesenchymal stem cells on microcarriers. This miniature auto-perfusion system provides an excellent solution to maintain cell-favorable conditions and high cell concentration in small-scale cultures for research and clinical uses.
中灌流对维持培养物中高细胞浓度至关重要。传统的膜过滤法在长期培养中面临膜过滤器堵塞和污染的挑战。在本研究中，我们提出了一种可在普通大小的实验室培养箱内操作的微型自动灌流系统。该系统配备了用于细胞保留的螺旋微流控芯片，以替代传统的膜过滤器，从根本上解决了堵塞和污染问题。我们表明，该系统支持悬浮培养的中国仓鼠卵巢（CHO）细胞连续灌注培养长达 14 天，无需更换细胞保留芯片。与每日手动换液相比，在 70ml 工作体积（2.8 VVD）的摇瓶中以 196ml/天的平均自动灌注率可维持 25%更高的 CHO 细胞浓度。自动灌流系统还在高浓度下产生了更好的细胞质量，表现为更高的活力、更均匀和规则的形态以及更少的聚集物。我们还证明了该系统在微载体上培养间充质干细胞的潜在应用。这种微型自动灌流系统为小规模培养中的细胞提供了极好的解决方案，可维持有利的条件和高细胞浓度，适用于研究和临床用途。
