CHO细胞无血清结团灌流培养：超声-沉降柱二合一灌流系统

利用CHO细胞能在培养过程中自然结团的特性，采用超声—沉降柱二合一灌流系统能促进细胞结团和加强截留的特性，我们用无血清培养基连续灌流培养基因重组CHO细胞MK3-A2株，分泌表达rhTNK-tPA获得了成功。培养周期为77-110天，细胞结团率为90%左右，直径在285～570μm之间，细胞截留率保持在95%左右，成活率为85%以上，细胞密度达到2×107/ml左右，rhTNK-tPA生产率平均为89 mg/L/d，最高时达216mg/L/d。 结果表明，使用该灌流系统进行细胞结团培养可以取代微载体培养用于动物细胞制药的规模化生产。

Serum free Aggregate Perfusion Culture of CHO Cells:an Ultrasonic and Sedimentation Column Combined Perfusion System

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor, we cultured successfully recombinant CHO cell strain MK3-A2, which could secrete TNK-tPA, by a serum-free perfusion culture system. The culture periods in our two experiments were as long as 77 and 110 days respectively. The cells density reached 2×107 cells /ml. The average volumetric productivity of TNK-tPA was 89 mg/L.d, and the highest one was 216mg/L.d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570μm. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.