Disposable 600-mL orbitally shaken bioreactor for mammalian cell cultivation in suspension

摘要

Orbitally shaken bioreactors (OSRs) support the efficient cultivation of mammalian cells in suspension at various scales of operation. Here we introduce a disposable culture vessel with a ventilated cap having a nominal volume of 600 mL (TubeSpin~R bioreactor 600) for medium-scale cell cultivation. It has a conical bottom designed to fit into a standard swinging bucket rotor to facilitate cell/liquid separation by centrifugation. At shaking frequencies suitable for mammalian cell cultivation (140-220rpm), the volumetric mass transfer coefficient (k_La) ranged from 15 to 40 h~(-1), the mixing time was less than 20 s, and the specific power consumption ranged from 0.03 to 0.5kW/m3. To define the working conditions for the TubeSpin~R bioreactor 600, we measured cell growth and environmental culture conditions such as pH and dissolved O2 and CO2 concentrations over a range of shaking frequencies and working volumes using a stable CHO-derived cell line expressing a human IgG antibody. The optimal cultivation conditions were observed at working volumes of 300-500mL and shaking frequencies of 180-220 rpm. In 300-mL batch cultures of the same cell line in a TubeSpin~R bioreactor 600,1-L cylindrical glass bottle, and 1-L Erlenmeyer shake flask, the maximum cell densities and recombinant antibody yields were similar. This study demonstrated the suitability of the TubeSpin~R bioreactor 600 for cell culture applications with suspension-adapted CHO cells at culture volumes up to 500 mL