Liu Xingmao, Liu Hong, Ye Lingling, Li Shichong, Wu Benchuan, Wang Haitao, Xie Jing, Chen Zhaolie
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Aug;26(8):1116-22.
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 (P < 0.05). The response surface methodology with three factors and three levels was used to determine the optimal levels of these factors. And a serum-free medium, SFM-CHO-S for recombinant CHO cells suspension culture was formulated. The maximum cell density of 11G-S cells in SFM-CHO-S in suspension batch culture reached 4.12 x 10(6) cells/mL with a maximum pro-UK activity at 5614 IU/mL, which was superior to the commercial serum-free medium for recombinant CHO cells.
以表达人尿激酶原（pro-UK）的悬浮适应型重组中国仓鼠卵巢（CHO）细胞系11G-S为研究对象，采用Plackett-Burman设计和响应面方法，配制了一种用于悬浮培养重组CHO细胞的无血清培养基。采用二级Plackett-Burman设计评估了10种培养基添加物对悬浮培养的11G-S细胞生长的影响。在这10种培养基添加物中，胰岛素、转铁蛋白和腐胺被确定为最显著的因素（P < 0.05）。采用三因素三水平的响应面方法确定这些因素的最佳水平。并配制了一种用于重组CHO细胞悬浮培养的无血清培养基SFM-CHO-S。在SFM-CHO-S中进行悬浮分批培养时，11G-S细胞的最大细胞密度达到4.12×10⁶个细胞/mL，最大尿激酶原活性为5614 IU/mL，优于重组CHO细胞的商业无血清培养基。
