Department of Food Science, Food and Bioprocess Engineering Group, Wageningen Agricultural University, P.O. Box 8129, 6700 EV, Wageningen, The Netherlands.
Cytotechnology. 1996 Jan;21(3):263-77. doi: 10.1007/BF00365349.
Hybridomas were cultured under steady-state conditions in a series of two continuous stirred-tank reactors (CSTRs), using a serum-free medium. The substrate not completely converted in the first CSTR, was transported with the cells to the second one and very low growth rates, high death rates, and lysis of viable cells were observed in this second CSTR. These conditions are hardly accessible in a single vessel, because such experiments would be extremely time-consuming and unstable due to a low viability. In contrast to what is often observed in literature, kinetic parameters could thus be derived without the neccessity for extrapolation to lower growth rates. Good agreement with literature averages for other hybridomas was found. Furthermore, showing that the reactor series is a valuable research tool for kinetic studies under extreme conditions, the possibility to observe cell death under stable and defined steady-state conditions offers interesting opportunities to investigate apoptosis and necrosis. Additionally, a model was developed that describes hybridoma growth and monoclonal antibody production in the bioreactor cascade on the basis of glutamine metabolism. Good agreement between the model and the experiments was found.
杂交瘤在一系列两个连续搅拌釜式反应器 (CSTR) 中在无血清培养基中于稳态条件下培养。在第一个 CSTR 中未完全转化的底物与细胞一起输送到第二个 CSTR 中，在第二个 CSTR 中观察到非常低的生长速率、高死亡率和活细胞裂解。由于生存力低，这种条件在单个容器中几乎无法实现，因为这样的实验将非常耗时且不稳定。与文献中经常观察到的情况相反，因此可以在无需推断至更低生长速率的情况下得出动力学参数。还发现与其他杂交瘤的文献平均值吻合良好。此外，该反应器系列证明是在极端条件下进行动力学研究的有价值的研究工具，能够在稳定和定义的稳态条件下观察细胞死亡，为研究细胞凋亡和坏死提供了有趣的机会。此外，还开发了一个模型，该模型基于谷氨酰胺代谢描述了生物反应器级联中杂交瘤的生长和单克隆抗体的生产。模型与实验之间存在良好的一致性。
