Rhiel M, Mitchell-Logean C M, Murhammer D W
Department of Chemical and Biochemical Engineering, The University of Iowa, 125B Chemistry Building, Iowa City, Iowa 52242, USA.
Biotechnol Bioeng. 1997 Sep 20;55(6):909-20. doi: 10.1002/(SICI)1097-0290(19970920)55:6<909::AID-BIT8>3.0.CO;2-K.
Nutrient utilization and byproduct accumulation were monitored in Spodoptera frugiperda Sf-9 and Trichoplusia ni BTI-Tn-5B1-4 (High Five) cell lines during growth and following viral infection in suspension cultures in order to develop a better understanding of cell metabolism and to acquire information relevant to large scale fed-batch bioreactors. The utilization of glucose, dissolved oxygen, and amino acids were monitored in Sf-9 cell cultures grown in Sf-900 II serum-free medium (SFM) and in High Five cell cultures grown in both Sf-900 II and Express Five SFM. Using the optimal medium for each cell line, i.e., Sf-900 II SFM for Sf-9 cells and Express Five SFM for High Five cells, the cell growth rate, maximum cell density, specific glucose and glutamine utilization rates, and specific alanine production rate were comparable during cell growth. In addition, the expression level of recombinant human tissue plasminogen activator was comparable in the two cell lines on a per cell basis. It was found, however, that lactate and ammonia accumulated in High Five cell cultures, but not in Sf-9 cell cultures. In addition, High Five cells utilized asparagine more rapidly than glutamine, whereas Sf-9 cells consumed only minimal asparagine, and the oxygen utilization rate was significantly higher in High Five cell cultures. It was also found that the medium had a significant effect on High Five cell metabolism, e.g., the specific glucose utilization rate and the specific lactate and alanine production rates were significantly higher in Sf-900 II SFM than in Express Five SFM. In addition, the maximum cell density and specific asparagine utilization rate were significantly higher in Express Five SFM.
为了更好地理解细胞代谢并获取与大规模补料分批生物反应器相关的信息，在悬浮培养中监测了草地贪夜蛾Sf-9和粉纹夜蛾BTI-Tn-5B1-4（High Five）细胞系在生长过程中和病毒感染后的营养物质利用及副产物积累情况。在无血清培养基Sf-900 II中生长的Sf-9细胞培养物以及在Sf-900 II和Express Five无血清培养基中生长的High Five细胞培养物中，监测了葡萄糖、溶解氧和氨基酸的利用情况。使用适合每个细胞系的最佳培养基，即Sf-9细胞用Sf-900 II无血清培养基，High Five细胞用Express Five无血清培养基，在细胞生长过程中，细胞生长速率、最大细胞密度、葡萄糖和谷氨酰胺的比利用率以及丙氨酸的比产生率相当。此外，重组人组织型纤溶酶原激活剂在两个细胞系中基于每个细胞的表达水平相当。然而，发现High Five细胞培养物中积累了乳酸和氨，而Sf-9细胞培养物中没有。此外，High Five细胞比谷氨酰胺更快地利用天冬酰胺，而Sf-9细胞仅消耗极少的天冬酰胺，并且High Five细胞培养物中的氧利用率明显更高。还发现培养基对High Five细胞代谢有显著影响，例如，Sf-900 II无血清培养基中的葡萄糖比利用率以及乳酸和丙氨酸的比产生率明显高于Express Five无血清培养基。此外，Express Five无血清培养基中的最大细胞密度和天冬酰胺比利用率明显更高。
