CHO工程细胞(11G-S)悬浮培养的无血清培养基的设计.PDFVIP

生 物 工 程 学 报 Chin J Biotech 2010, August 25; 26(8): 1116−1122 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ ©2010 CJB, All rights reserved. 组织工程与细胞培养 CHO 工程细胞 (11G-S) 悬浮培养的无血清培养基的 设计 刘兴茂，刘红，叶玲玲，李世崇，吴本传，王海涛，谢靖，陈昭烈 军事医学科学院生物工程研究所，北京 100071 摘 要: 以悬浮适应的表达重组尿激酶原 (Pro-urokinase ，pro-UK) CHO 工程细胞系11G-S 为对象，采用Plackett-Burman 实验设计及响应面分析法，设计支持 CHO 工程细胞 (11G-S) 悬浮生长的无血清培养基。以细胞密度为评价指标，在 单因素实验的基础上采用Plackett-Burman 实验设计对影响细胞生长的培养基添加成分进行考察，确定了3 种对细胞生 长明显促进作用的培养基添加成分：胰岛素、转铁蛋白及腐胺。继而利用响应面法分析了这 3 种添加成分的最佳水平 范围，设计了一种适用于 CHO 工程细胞 (11G-S) 悬浮培养的无血清培养基 SFM-CHO-S。11G-S 细胞在 SFM-CHO-S 批次悬浮培养的细胞最大生长密度达到4.12×106 cells/mL ，pro-UK 的最大累积活性达到5 614 IU/mL ，培养效果优于商 品化的同类无血清培养基。 关键词: CHO 工程细胞，悬浮培养，无血清培养基，实验设计 Serum-free medium for suspension culture of recombinant Chinese hamster ovary (11G-S) cells Xingmao Liu, Hong Liu, Lingling Ye, Shichong Li, Benchuan Wu, Haitao Wang, Jing Xie, and Zhaolie Chen Institute of Biotechnology , Academy of Military Medical Sciences, Beijing 100071, China Abstract: With suspension adapted recombinant Chinese hamster ovary (CHO) cell lines 11G-S expressing human pro-urokinase (pro-UK) as the object of study, a serum-free medium for the cultivation of recombinant CHO cells in suspension was formulated by using Plackett-Burman design and response surface methodology. The two-level Plackett-Burman design was used to evaluate the effect of 10 medium supplements on the growth of the 11G-S cells in suspension culture. Among the 10 medium supplements, insulin, transferrin, and putrescine were identified as the most significant factors (P0.05). The response surface methodology with three factors and three levels was used to determine the optimal lev