Ozturk S S, Palsson B O
Department of Chemical Engineering, University of Michigan, Ann Arbor 48109.
Biotechnol Prog. 1991 Nov-Dec;7(6):471-80. doi: 10.1021/bp00012a001.
A mouse-mouse hybridoma cell line (167.4G5.3) was cultivated in a 1.5-L stirred-tank bioreactor under constant pH and dissolved oxygen concentration. The transient kinetics of cell growth, metabolism, and antibody production were followed by biochemical and flow cytometric methods. The cell-specific kinetic parameters (growth and metabolic rates) as well as cell size were constant throughout the exponential phase. Intracellular protein and RNA content followed a similar trend. Cell growth stopped when the glutamine in the medium was depleted. Glucose could not substitute for glutamine, as glucose consumption ceased after glutamine depletion. Ammonia and lactate production followed closely glutamine and glucose consumption, respectively. Alanine, glutamate, serine, and glycine were produced but other amino acids were consumed. The cells are estimated to obtain about 45% of the total energy from glycolysis, with the balance of the metabolic energy provided by oxidative phosphorylation. The antibody was produced at a constant rate in both the exponential and decline phases of growth. The intracellular antibody content of the cells remained relatively constant during the exponential phase of growth and decreased slightly afterwards.
将小鼠-小鼠杂交瘤细胞系(167.4G5.3)在1.5升搅拌罐生物反应器中于恒定pH值和溶解氧浓度下培养。通过生化和流式细胞术方法跟踪细胞生长、代谢和抗体产生的瞬态动力学。在整个指数生长期,细胞特异性动力学参数(生长和代谢速率)以及细胞大小保持恒定。细胞内蛋白质和RNA含量呈现相似趋势。当培养基中的谷氨酰胺耗尽时,细胞生长停止。葡萄糖不能替代谷氨酰胺,因为谷氨酰胺耗尽后葡萄糖消耗停止。氨和乳酸的产生分别与谷氨酰胺和葡萄糖的消耗密切相关。细胞产生了丙氨酸、谷氨酸、丝氨酸和甘氨酸,但消耗了其他氨基酸。据估计,细胞约45%的总能量来自糖酵解,其余代谢能量由氧化磷酸化提供。在生长的指数期和衰退期,抗体均以恒定速率产生。在生长的指数期,细胞内抗体含量保持相对恒定,之后略有下降。