Cultivation of Recombinant Chinese Hamster Ovary Cells Grown as Suspended Aggregates in Stirred Vessels(BIOCHEMICAL ENGINEERING)

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概要

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• Recombinant Chinese hamster ovary (rCHO) cells capable of producing a prourokinase mutant (mPro-uk) grown as suspended aggregates in stirred vessels were described and characterized. The addition of chitosan to a mixture of DMEM and Ham's F12 (D-MEM/F-12) medium promoted cell aggregation and spheroid formation efficiently. Multicellular aggregates formed immediately after the rCHO cells were inoculated into the chitosan-added medium, and the mean diameter of the cell aggregates reflecting the aggregate size increased with culture time, shifting from 65 to 163μm after 2 and 9d of culture in spinner flasks. No significant difference in the metabolism performance of the rCHO cells was observed between suspended aggregates and anchored monolayers. However, the cells cultured as suspended aggregates showed a marked decrease in growth rate as evaluated from specific growth rate (μ). Replacing D-MEM/F-12 medium with CD 293 medium caused compact spherical cell aggregates to dissociate into small irregular aggregates and single cells without apparent effects on cell performance in subcultures. The perfusion culture of the rCHO cells grown as suspended aggregates in a 2-l stirred tank bioreactor for 15d resulted in a maximum viable cell density of 5.6×10^6 cells ml^<-1> and an mPro-uk concentration of about 2.6×10^3 IU ml^<-1>, and cell viability was remained at roughly 90% during the entire run.

• 社団法人日本生物工学会の論文
• 2006-11-25

著者

• LIU Hong

  Department of Orthodontics, School of Stomatology, Shanghai Second Medical University

• Liu Xing-Mao

  Department of Cell Engineering, Beijing Institute of Biotechnology

• Li Shi-Chong

  Department of Cell Engineering, Beijing Institute of Biotechnology

• Wu Ben-Chuan

  Department of Cell Engineering, Beijing Institute of Biotechnology

• Ye Ling-Ling

  Department of Cell Engineering, Beijing Institute of Biotechnology

• Wang Qi-Wei

  Department of Cell Engineering, Beijing Institute of Biotechnology

• Chen Zhao-Lie

  Department of Cell Engineering, Beijing Institute of Biotechnology

• HAN Yi

  Department of Applied Chemistry, Faculty of Engineering, The University of Tokyo

• Liu Hong

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Wang Qu-wei

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Wang Qi-wei

  Dep. Of Cell Engineering Beijing Inst. Of Biotechnology 20 Dongdajie Street Fengtai Beijing 100071 C

• Ye Ling-ling

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Liu Xing-mao

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Wu Ben-chuan

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Li Shi-chong

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Chen Zhao-lie

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Han Yi

  Department Of Cell Engineering Beijing Institute Of Biotechnology

• Han Yi

  Department Of Applied Chemistry Graduate School Of Engineering The University Of Tokyo

• Liu Hong

  Department Of Anesthesiology And Pain Medicine University Of California Davis Medical Center

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