Kurano N, Leist C, Messi F, Kurano S, Fiechter A
Department of Biotechnology, Swiss Federal Institute of Technology, Zürich.
J Biotechnol. 1990 Jul;15(1-2):101-111. doi: 10.1016/0168-1656(90)90054-f.
Chinese hamster ovary (CHO) cells were cultivated in a compact loop bioreactor using MEM-alpha medium supplemented with 10% fetal calf serum. Effects of physical and chemical environments, i.e., pH in the medium, stirring speed of impellers, temperature and partial pressure of oxygen (pO2) upon growth of suspended cells in the bioreactor were determined in batch cultures. Growth behavior was characterized by specific rates of growth (mu), glucose consumption (qG) and lactate production (qL), and the yield coefficients (cell yield from glucose, YX/G, and lactate yield from glucose, YL/G). An effect of medium osmolality was also evaluated with T-flask monolayer cultivation. The best growth was observed at pH 7.6, 37 degrees C, 400 rpm, 50-100% saturation with oxygen and 320 mOsmol kg-1. Corresponding to the previous work with a human melanoma cell line, the sophisticated cultivation and process control systems have been improved for CHO cells.
中国仓鼠卵巢(CHO)细胞在紧凑型环路生物反应器中培养,使用添加了10%胎牛血清的MEM-α培养基。在分批培养中,测定了物理和化学环境,即培养基中的pH值、搅拌桨的搅拌速度、温度和氧分压(pO2)对生物反应器中悬浮细胞生长的影响。生长行为通过特定生长速率(μ)、葡萄糖消耗速率(qG)和乳酸产生速率(qL)以及产率系数(基于葡萄糖的细胞产率,YX/G,和基于葡萄糖的乳酸产率,YL/G)来表征。还通过T型瓶单层培养评估了培养基渗透压的影响。在pH 7.6、37℃、400 rpm、50-100%氧气饱和度和320 mOsmol kg-1条件下观察到最佳生长。与之前对人黑色素瘤细胞系的研究工作一致,针对CHO细胞改进了复杂的培养和过程控制系统。