Ozturk S S, Palsson B O
Department of Chemical Engineering, University of Michigan, Ann Arbor 48109.
Biotechnol Prog. 1990 Nov-Dec;6(6):437-46. doi: 10.1021/bp00006a006.
The effects of dissolved oxygen concentration (DO) on hybridoma cell physiology were examined in a continuous stirred tank bioreactor with a murine hybridoma cell line (167.4G5.3). Dissolved oxygen concentration was varied between 0% and 100% air saturation. Cell growth and viability, carbohydrate, amino acid, and energy metabolism, oxygen uptake, and antibody production rates were investigated. Cell growth was inhibited at both high and low DO. Cells could grow at 0% DO and maintain viability under a nitrogen atmosphere. Cell viability was higher at low DO. Glucose, glutamine, and oxygen consumption rates changed little at DO above 1% air saturation. However, the metabolic uptake rates changed below 1% DO, where growth became oxygen limited, and a Km value of 0.6% DO was obtained for the specific oxygen uptake rate. The metabolic rates of glucose, glutamine, lactate, and ammonia increased 2-3-fold as the DO dropped from 1% to 0%. Amino acid metabolism followed the same general pattern as that of glutamine and glucose. Alanine was the only amino acid produced. The consumption rates of amino acids changed little above 1% DO, but under anaerobic conditions the consumption rates of all amino acids increased severalfold. Cells obtained most of their metabolic energy from glutamine oxidation except under oxygen limitation, when glucose provided most of the energy. The calculated ATP production rate was only slightly influenced by DO and rose at 0% DO. Antibody concentration was highest at 35% DO, while the specific antibody production rate was insensitive to DO.
在一个连续搅拌罐式生物反应器中，使用鼠杂交瘤细胞系（167.4G5.3）研究了溶解氧浓度（DO）对杂交瘤细胞生理学的影响。溶解氧浓度在0%至100%空气饱和度之间变化。研究了细胞生长和活力、碳水化合物、氨基酸和能量代谢、氧气摄取以及抗体产生速率。高溶解氧和低溶解氧均抑制细胞生长。细胞可以在0%溶解氧下生长，并在氮气气氛中维持活力。低溶解氧时细胞活力更高。在溶解氧高于1%空气饱和度时，葡萄糖、谷氨酰胺和氧气消耗速率变化不大。然而，在溶解氧低于1%时，代谢摄取速率发生变化，此时生长受到氧气限制，特定氧气摄取速率的Km值为0.6%溶解氧。随着溶解氧从1%降至0%，葡萄糖、谷氨酰胺、乳酸和氨的代谢速率增加了2至3倍。氨基酸代谢遵循与谷氨酰胺和葡萄糖相同的一般模式。丙氨酸是唯一产生的氨基酸。在溶解氧高于1%时，氨基酸消耗速率变化不大，但在厌氧条件下，所有氨基酸的消耗速率均增加了几倍。除了在氧气限制条件下葡萄糖提供大部分能量外，细胞的大部分代谢能量来自谷氨酰胺氧化。计算得出的ATP产生速率仅受溶解氧的轻微影响，并在0%溶解氧时上升。抗体浓度在35%溶解氧时最高，而特定抗体产生速率对溶解氧不敏感。
