Abstract
accumulation, high mixing times, poorair dispersion and concentration gradients. In batchreactor, we have previously found that growth kineticsof CHO cells could be enhanced by increasing stirringfrequencies from 80 to 600 rpm. However, these studieswere limited in time due to depletion of substrates lead-ing to a cell death which is not associated to hydrody-namic constraints. To overcome the effects on celldeath from those nutritional limitations, in this work,longer stationary cultures of CHO cells were carried outin continuous mode in a 2-liter reactor operating withvarious stirring profiles.Materials and methodsCHO 320 cells were grown in a serum-free medium PF-BDM in a sparged and stirred tank reactor (1.4 L work-ing volume; pO
Highlights
Mammalian cells are known to be sensitive to hydrodynamic stresses
Continuous culture performed with the first agitation rate profile : 300-600-300-600 rpm Based on previous batch culture results [1], the batch phase was started at 300 rpm during 48 h in order to obtain the mid-exponential cell growth phase before starting the continuous mode (Figure 1A)
Viable cell concentration increased to 30.105 cells.mL-1 at the steady state, while dead and lysed cells reached an average level of 5.105 cells.mL-1
Summary
Mammalian cells are known to be sensitive to hydrodynamic stresses. Soft agitation and aeration are generally recommended in culture bioreactor to prevent cell damages. We have previously found that growth kinetics of CHO cells could be enhanced by increasing stirring frequencies from 80 to 600 rpm. These studies were limited in time due to depletion of substrates leading to a cell death which is not associated to hydrodynamic constraints. To overcome the effects on cell death from those nutritional limitations, in this work, longer stationary cultures of CHO cells were carried out in continuous mode in a 2-liter reactor operating with various stirring profiles
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