Center for Advanced Sensor Technology, and Department of Chemical and Biochemical Engineering, University of Maryland-Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA.
Biotechnol Bioeng. 2010 Apr 15;105(6):1040-7. doi: 10.1002/bit.22649.
Routine cell culture is done in small-scale disposable vessels (typically 0.1-100 mL volumes) in academia and industry. Despite their wide use in bioprocess development (i.e., process optimization and process validation), miniature process scouting devices (PSDs) are considered "black boxes" because they are generally not equipped with sensors. In this study, we show that on-line monitoring of dissolved oxygen (DO) and pH in a T-75 flask-based PSD can be achieved during cell passaging and that this information can be linked to different cellular metabolic states. In this case, on-line monitoring of DO and pH show three distinctive metabolic regions in passages 1-18, 19-28, 29-54 and in particular, the shift in the pH curve, the specific oxygen uptake rate (q(O2)), and the lactate production rate to the oxygen consumption rate yield (Y(Lac/ox)) confirm the existence of these distinctive metabolic regions. These findings are particularly useful because they show that sensor equipped PSDs can help to monitor cell culture behavior after thaw, in pre- and seed culture prior to scale-up and in development/optimization studies. Such routine monitoring will help to develop more consistent cell culture techniques.
在学术界和工业界中，常规的细胞培养是在小规模的一次性容器（通常为 0.1-100 毫升体积）中进行的。尽管微型过程探测设备（PSD）在生物过程开发（即过程优化和过程验证）中被广泛使用，但由于它们通常没有配备传感器，因此被认为是“黑匣子”。在本研究中，我们表明，在细胞传代过程中，可以在线监测 T-75 瓶式 PSD 中的溶解氧（DO）和 pH 值，并且可以将这些信息与不同的细胞代谢状态相关联。在这种情况下，DO 和 pH 值的在线监测显示出在传代 1-18、19-28、29-54 过程中的三个不同的代谢区域，特别是 pH 值曲线、特定耗氧速率（q（O2））和乳酸生成速率与氧消耗速率之比（Y（Lac/ox））的变化证实了这些不同代谢区域的存在。这些发现非常有用，因为它们表明配备传感器的 PSD 可以帮助在解冻后、在放大前的预培养和种子培养以及在开发/优化研究中监测细胞培养行为。这种常规监测将有助于开发更一致的细胞培养技术。
