昆虫细胞(Sf21)悬浮培养过程中生长限制性基质间歇补加技术的应用

基于Sf21昆虫细胞在悬浮培养过程中所表现出的生长代谢特征，提出以培养液中残糖浓度作为控制参数，并利用限制性基质(葡萄糖和蛋白水解物)的间歇补加技术调控细胞生长的方案。实际控制表明：与批培养相比，Sf21细胞在两种具代表性的昆虫细胞培养基(IPL-41和TC-100)中的生长期和稳定期都得到了有效的延长。TC-100培养液中最高细胞培养密度由3.0×10⁶ cells/mL提高到6.5×10⁶ cells/mL；IPL41培养液中最高细胞培养密度则由7.05×10⁶ cells/mL提高到9.0×10⁶cells/mL。由于限制性基质的间歇补加技术是利用较确定的营养成分来代替复杂昂贵的补料培养基，因此更适合于昆虫细胞的大规模高密度培养。

Application of Intermittent-feeding of Growth-limiting Nutrients in Suspension Culture of Insect Cells(Sf21)

On the basis of the growth and metabolism behavior inherent in suspended Sf21 insect cells,the intermittent feeding of growth-limiting nutrients(glucose and protein hydrolysates)was employed in the regulation of cellular growth during the cultivation by using the residual glucose concentration as a reference point. It was shown that as compared with the batch cultivation, the intermittent-feeding of growth-limiting nutrients effectively prolonged the growth and stationary phase for Sf21 insect cells grown in two representative culture medium(TC-100 and IPL-41). The maximum cell density was increased from 3.0×106 cells/ml to 6.5×106 cells/mL in TC-100 medium, and in IPL-41 medium, the maximum cell density was increased from 7.05×106 cells/mL to 9.0×106 cells/mL. As the defined nutrient solution was used for feeding in lieu of the complicated and expensive base medium, the technique would find prospects in large scale high-density cultivation of insect cells.