High density culture of insect cells using rational medium design and feeding strategy
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : T.-W. Chiou, Y.-C. Hsieh, C. S. Ho
- چاپ و سال / کشور: 2000
Description
The objective of this study is to achieve high density cell culture by a rational medium design and feeding strategy. Insect cell/baculovirus expression system is one of the widely used methods for the production of heterologous proteins in the cell culture domain. Insect cell Spodoptera frugiperda Sf-21 and a recombinant baculovirus with encoded gene for human interleukin-5 were chosen as the model system in this study. A stoichiometric model was established to study the demand of nutrients, including glucose, 20 amino acids, and yeastolate, for the synthesis of cell mass. The coef®cients for individual nutrients in the stoichiometric equation governing insect cell growth were determined from the information of cell mass and compositions. Based on the stoichiometric coef®- cients, the initial and supplemental media for fed-batch cell cultures were designed. The experiments began with the inoculation of Sf-21 cells into a spinner ¯ask with the initial medium, which provided a starting environment for achieving optimum cell growth. This was followed by the periodic feeding of supplemental medium designed by utilizing the stoichiometric equation that governs insect cell growth. With this strategy, it was demonstrated that the Sf-21 cell culture reached a cell density in excess of 1:9 107 cells/ml. During the cultivation process, the utilization of various nutrients and the production of metabolites were also monitored. Further experiments proved that high concentration of recombinant product (such as human interleukin- 5) could be achieved by infecting the high density cells (resulting from the designed medium) with recombinant baculoviruses.
