標題: High density culture of insect cells using rational medium design and feeding strategy
作者: Chiou, TW
Hsieh, YC
Ho, CS
生物科技學系
Department of Biological Science and Technology
公開日期: 1-Jun-2000
摘要: 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 coefficients 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 coefficients, 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 flask 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 x 10(7) 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.
URI: http://hdl.handle.net/11536/30488
ISSN: 0178-515X
期刊: BIOPROCESS ENGINEERING
Volume: 22
Issue: 6
起始頁: 483
結束頁: 491
Appears in Collections:Articles


Files in This Item:

  1. 000088149100003.pdf