Stavroulakis D A, Kalogerakis N, Behie L A, Iatrou K
Pharmaceutical Production Research Facility (PPRF), The University of Calgary, Calgary, Alberta T2N 1N4.
Biotechnol Bioeng. 1991 Jun 20;38(2):116-26. doi: 10.1002/bit.260380203.
The kinetics and growth characteristics of the BM-5 insect cell line of Bombyx mori (silkworm) have been experimentally investigated in order to develop optimal growth protocols when these cells are used to produce large quantities of human proteins by recombinant baculoviruses. Experiments were performed in 200-mL spinner flasks, which were operated at 80 rpm with 0.3% methyl cellulose (MCL) added to the medium to protect the cells from shear stress. Exposure of the cells to varying nutrient and metabolite concentrations was accomplished through a repeated-batch mode of feeding. The results indicate that glutamine is a limiting nutrient and that lactate has an inhibitory effect on cell growth. Ammonia depletion from the medium was accompanied by uric acid accumulation suggesting that ammonia is converted to this metabolic product by the "uricotelic" and "nucleicolytic" metabolic pathways. With the examined medium formulation, glucose, fructose, and sucrose remained at high concentrations throughout the cultivation period.
为了制定最佳生长方案，以便利用家蚕BM-5昆虫细胞系通过重组杆状病毒大量生产人类蛋白质，对其动力学和生长特性进行了实验研究。实验在200毫升旋转瓶中进行，转速为80转/分钟，培养基中添加0.3%的甲基纤维素（MCL）以保护细胞免受剪切应力。通过重复分批补料方式使细胞暴露于不同的营养物和代谢物浓度下。结果表明，谷氨酰胺是一种限制性营养物质，乳酸对细胞生长具有抑制作用。培养基中氨的消耗伴随着尿酸的积累，这表明氨通过“排尿酸”和“核酸分解”代谢途径转化为这种代谢产物。在所研究的培养基配方中，葡萄糖、果糖和蔗糖在整个培养期间都保持在高浓度。
