Abstract：
Cell line generation of mammalian cells is a time-consuming and labor-intensive process, especially because of challenges in clone selection after transfection. Antibiotics are common selection agents for mammalian cells due to their simplicity of use. However, the optimal antibiotic concentration must be determined with a kill curve experiment before clone selection starts. The traditional kill curve experiments are resource-intensive and time-consuming due to necessary sampling and offline analysis effort. This study, thus, explores the potential of online monitoring the oxygen transfer rate (OTR), as a non-invasive and efficient alternative for kill curve experiments. The OTR is monitored using the Transfer-rate Online Measurement (TOM) system and the micro(μ)-scale Transfer-rate Online Measurement (μTOM) device, which was used for mammalian cells first. It could be shown that the OTR curves for both devices align perfectly, affirming consistent cultivation conditions. The μTOM device proves effective in performing kill curve experiments in 96-deep-well plates without the need for sampling and offline analysis. The streamlined approach reduces medium consumption by 95%, offering a cost-effective and time-efficient solution for kill curve experiments. The study validates the generalizability of the method by applying it to two different CHO cell lines (CHO-K1 and sciCHO) with two antibiotics (puromycin and hygromycin B) each. In conclusion, the broad application of OTR online monitoring for CHO cell cultures in 96-deep-well plates is highlighted. The μTOM device proves as a valuable tool for high-throughput experiments, paving the way for diverse applications, such as media and clone screening, cytotoxicity tests, and scale-up experiments.
摘要：
哺乳动物细胞的细胞系生成是一个耗时耗力的过程,特别是由于转染后克隆选择的挑战。抗生素由于其使用简单而成为哺乳动物细胞的常用选择剂。然而,在克隆选择开始之前，必须通过杀死曲线实验确定最佳抗生素浓度。由于必要的采样和离线分析工作，传统的杀灭曲线实验资源密集且耗时。这项研究,因此，探索了在线监测氧气传输速率(OTR)的潜力，作为杀死曲线实验的非侵入性和有效的替代方法。使用传输速率在线测量(TOM)系统和微(μ)尺度传输速率在线测量(μTOM)装置监测OTR,首先用于哺乳动物细胞。可以看出，两种设备的OTR曲线完全对齐，确认一致的栽培条件。μTOM装置证明可有效地在96深孔板中进行杀灭曲线实验，而无需采样和离线分析。精简的方法减少了95%的介质消耗，为杀伤曲线实验提供具有成本效益和时间效率的解决方案。该研究通过将其应用于两种不同的CHO细胞系（CHO-K1和sciCHO），并分别使用两种抗生素（嘌呤霉素和潮霉素B）来验证该方法的通用性。总之,强调了OTR在线监测在96深孔板中CHO细胞培养物中的广泛应用。μTOM设备被证明是高通量实验的宝贵工具，为不同的应用铺平道路，如培养基和克隆筛选，细胞毒性试验，和放大实验。
