Abstract:

Cell culture technology is the pillar of the biopharmaceutical industry. T-flasks as single-use consumables are typically used in single-layer static cultivation which produces limited cell density. In order to achieve high-throughput and high density cell culture in T-flasks, this study systematically investigates the hydrodynamics and mass transfer characteristics of a T25 flask placed on a rocking platform under different operating conditions. The results show that shaking can significantly increase the mass transfer rate of the square bottle and reduce the mixing time, making high-density culture possible. But the air filter on the bottle cap becomes the limiting factor for the mass transfer rate at high rotation speeds. The horizontal and vertical distances of the flask relative to the rocking axis had negligible effects on mass transfer and mixing, but placing the flask at a 45° angle and effectively using the walls of the flask as baffles, drastically reduces the mixing time at the same rocking speed. A user-defined function is used to implement a dynamic mesh to simulate the rocking movement of the T-flask in a CFD model. The CFD model enables the analysis of the shear stress and energy dissipation rates in the flask under different rocking speeds. This study provides the data and theoretical support for the further development of a T-flask-based high-throughput, single-use microbioreactor systems.

Key words: T25 flask, cell culture, mixing, mass transfer, CFD

摘要：

细胞培养技术是生物医药产业的支柱，以实现微小体积内的高密度、高通量细胞培养为目的，系统地研究了常用于单层静态培养的T25方瓶置于翘板摇床上在不同操作条件下的流体力学特性和传质性能。结果表明，振荡可以显著提高方瓶的传质速率并降低混合时间，使高密度培养成为可能，但瓶盖上的空气滤膜在高转速时成为传质速率的限制因素；培养瓶对称轴与摇床旋转轴平行时，其相对位置对混合和传质无明显影响，但当二者成45°角时，相同转速下混合时间显著缩短；使用自定义函数实现了基于动态网格的CFD模拟，对不同转速下方瓶内剪切应力和能量耗散在时间与空间上分布进行了分析，为基于T25培养瓶开发一次性高通量微型反应器提供了数据支持和理论基础。

关键词: T25方瓶, 细胞培养, 混合, 传质, 计算流体力学