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›› 2011, Vol. 11 ›› Issue (6): 1050-1054.

• 生化工程专栏 • 上一篇    下一篇

利用新型一次性激流灌注式生物反应器培养动物细胞

谢波 李峦峰 杜春玲 郝素娟 吴书音 惠觅宙   

  1. 中国科学院过程工程研究所生化工程国家重点实验室 杭州安普生物工程有限公司 杭州安普生物工程有限公司 中国科学院过程工程研究所生化工程国家重点实验室 杭州安普生物工程有限公司 中国科学院过程工程研究所生化工程国家重点实验室
  • 收稿日期:2011-09-20 修回日期:2011-11-18 出版日期:2011-12-20 发布日期:2011-12-20
  • 通讯作者: 谢波

Scale-up Cultivation of Animal Cells in a Novel Disposable Perfusion and Current Bioreactor

XIE Bo LI Luan-feng DU Chun-ling HAO Su-juan WU Shu-yin HUI Mi-zhou   

  1. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences AmProtein Co. Ltd AmProtein Co. Ltd National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences AmProtein Co. Ltd National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences
  • Received:2011-09-20 Revised:2011-11-18 Online:2011-12-20 Published:2011-12-20
  • Contact: XIE Bo

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被引次数

6

摘要： 利用新型一次性培养袋在5~10 L激流灌注式生物反应器中培养3种贴壁和3种悬浮细胞，通过在线控制培养温度、pH值和溶氧等工艺参数，监测并分析培养过程中细胞密度、活率、糖耗等的变化情况. 结果表明，贴壁培养细胞MDCK, VERO和DF-1培养6 d后消化计数，密度分别达4.8×106,1.0×107和1.5×107 mL-1，细胞状态良好，活率在90%以上. 悬浮细胞CHO, BHK-21和Sf9经无血清悬浮驯化后，经种子链扩增，在激流式反应器中悬浮培养的初始接种密度分别为2.0×106, 2.1×106和2.15×106 mL-1时，培养一段时间后其最高培养密度分别可达2.0×107, 2.1×107和1.8×107 mL-1，维持培养时间分别为13, 12和10 d.

关键词: 动物细胞, 培养, 贴壁, 悬浮, 一次性培养袋, 激流灌注式生物反应器

Abstract: A novel disposable perfusion and current bioreactor with 5~10 L work volume was used for scale-up cultivation of six kinds of animal cells. Several important parameters were controlled on-line, including cultivation temperature, pH value and dissolved oxygen (DO). The cell density, viability and consumption of glucose were determined and analyzed in the cultivation process. The concentration of adherent MDCK, VERO and DF-1 cells reached 4.8×106, 1.0×107 and 1.5×107 mL-1 after 6 d cultivation, respectively. They were in good condition and their viability was above 90%. After serum-free suspending domestication, the suspension cells of CHO, BHK-21 and Sf9 were transferred into the current bioreactor with the initial inoculation concentration of 2.0×106, 2.1×106 and 2.15×106 mL-1, respectively. The highest concentration of these cells reached about 2.0×107, 2.1×107 and 1.8×107 mL-1, the continuous cultivation time lasted 13, 12 and 10 d, respectively.

Key words: animal cell, cultivation, adherent, suspension, disposable bag, perfusion and current bioreactor

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