Zhang Weijian, Ran Qingyuan, Zhou Yang, Zhao Liang, Ye Qian, Tan Wen-Song
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, P. O. Box 309#, Shanghai, 200237 China.
Cytotechnology. 2025 Feb;77(1):7. doi: 10.1007/s10616-024-00662-x. Epub 2024 Nov 27.
Shake tubes (ST) are widely employed to assist the development of the stirred tank reactor (STR) perfusion cell culture. However, cell lysis may be frequently underrestimated and lead to culture performance discrepency between these systems, rendering the ST model ineffective in designing the STR perfusion cultures. In this study, perfusion culture performance bewteen the STR and ST was investigated under various conditions with the analysis of cell lysis. Comparable performance was observed bewteen the two systems at low perfusion rates ( ≤1.0 VVD), except that the specific productivity ( ) of the STR was decreased at =0.5 VVD, which was related to product degradation by cell lysis. In contrast, significant differences in cell maintenance, metabolism, and were found at =2.0 VVD. By the analysis of the authentic cell growth and death kinetics, it was found that cell growth arrest, potentially due to the limited availability of oxygen, led to the stable cell maintenance at VCD≈90 × 10 cells/ml and altered cellular metabolism for the ST, while the continuous decline of VCD and in the STR were related to excessive cell death, subsequently ascribed to the harmful hydrodynamic stress conditions. We further demonstrated that cell lysis accounted for 57.62-76.29% of the total generated biomass in both the reactors and significantly impacted the estimation of process descriptors crucial for understanding the true cellular states. With cell lysis in sight, cell performance can therefore be accurately described and this knowledge can be further leveraged to expedite process development for the perfusion cell culture processes.
摇瓶（ST）被广泛用于辅助搅拌罐反应器（STR）灌注细胞培养的开发。然而，细胞裂解情况可能经常被低估，从而导致这些系统之间的培养性能出现差异，使得ST模型在设计STR灌注培养时无效。在本研究中，通过分析细胞裂解，研究了STR和ST在各种条件下的灌注培养性能。在低灌注速率（≤1.0 VVD）下，观察到两个系统具有可比的性能，但在VVD = 0.5时，STR的比生产率（ ）有所下降，这与细胞裂解导致的产物降解有关。相比之下，在VVD = 2.0时，发现细胞维持、代谢和 存在显著差异。通过对真实细胞生长和死亡动力学的分析，发现细胞生长停滞可能是由于氧气供应有限，导致ST在VCD≈90×10细胞/ml时细胞维持稳定，并改变了细胞代谢，而STR中VCD和 的持续下降与过度细胞死亡有关，这随后归因于有害的流体动力学应力条件。我们进一步证明，细胞裂解在两个反应器中占总生成生物量的57.62 - 76.29%，并显著影响对理解真实细胞状态至关重要的过程描述符的估计。鉴于细胞裂解情况，因此可以准确描述细胞性能，并且可以进一步利用这些知识来加速灌注细胞培养过程的工艺开发。
