中国仓鼠卵巢细胞的悬浮适应及代谢特征

Suspension Adaptation and Metabolism of Chinese Hamster Ovary Cells

目的:通过悬浮适应,使中国仓鼠卵巢细胞(CHO细胞)获得悬浮生长的特性,并可在悬浮培养条件下较快地生长。方法:将CHO细胞以3×105/mL接种于100mL的三角瓶内,培养时加入1%小牛血清、1g/LPluronic F-68、25μg/mL硫酸葡聚糖,培养体积35mL,摇床转速90r/min,每24h离心换液,当细胞增殖为2×106/mL时传代。结果:经过悬浮适应,细胞的平均比生长速率由适应最初的0.27/d提高为适应后的0.48/d,最大总细胞密度由适应初期的2.5×106/mL提高为适应后的6.3×106/mL,目的蛋白活性也由适应前的2781U/mL提高为适应后的8878U/mL,适应后细胞的葡萄糖平均比消耗率为1.42μmol/(106细胞·d),低于适应前的2.16μmol/(106细胞·d)。结论:贴壁生长的CHO细胞经过悬浮适应,不仅可以在悬浮培养条件下快速生长,而且细胞对葡萄糖的利用率也得到提高。

Objective： Through suspension adaptation, to endow the Chinese hamster ovary cells（CHO cells） with suspension growth characteristics and rapid growth ability. Methods： Suspension culture was carried out in 100 mL shaker flasks with 35 mL culture volume and shaking at 90 r/min at an initial inoculation density of 3×10^5 cells/mL. The culture medium was supplemented with 1% serum, 1 g/L Pluronic F-68 and 25 μg/mL dextran sulfate. When the total cell density exceeded 2×10^6 cells/mL, the cells were subcuhured to 3×10^5 cells/mL. Results： Compared with the original cells, the average specific growth rate of the cells adapted to suspension culture increased from 0.27/d to 0.48/d, the highest density of total cells increased from 2.5×10^6 cells/mL to 6.3×10^6 cells/mL, and the activity of interest protein increased from the 2 781 U/mL to 8 878 U/mL. The average specific consumption rate of glucose of cells adapted to suspension culture was 1.42 μmol/（10^6 cells.d）, which was lower than that of the original cells [2.16 μmol/（10^6 cells＇d）]. Conclu-sion： The adherent CHO cells lines adapted to suspension culture can not only grow rapidly under suspension culture conditions, but also improve utilization efficiency to glucose of the cells.