High cell density suspension culture of mammalian anchorage independent cells: oxygen transfer by gas sparging and defoaming with a hydrophobic net.
Gas sparging directly into the culture-broth is not done in cell culture, except when the gas flow rate is very small, because much foaming occurs. During screening of defoaming methods, foam was observed to be broken up effectively when it made contact with a net fabricated from hydrophobic materials. Providing a highly efficient oxygen supply to suspension culture was tried using the new defoaming method. In a 5 l reactor equipped with the foam-eliminating net fabricated with polysiloxane, oxygen was transferred at 21 mmole/l.h equivalent to a consumption rate of 1 X 10(8) cells/ml, even at a low oxygen gas flow rate of 0.1 cm/s corresponding to a fourth of the gas flow rate when foam leaked through the net. Perfusion culture of rat ascites hepatoma cell JTC-1 was successfully carried out in the 5 l scale culture system with the net and a hydrophobic membrane for cell filtration. The viable cell concentration reached 2.7 X 10(7) cells/ml after twenty-seven days, in spite of the nutrient-deficient condition of the lower medium exchange rate, that is, a working volume a day, and viability was maintained at more than 90%. In a 1.21 scale culture of mouse-mouse hybridoma cell STK-1, viable cell concentration reached 4 X 10(7) cells/ml. These results showed that oxygen transfer by gas sparging with defoaming was useful for high density suspension culture. A foam-breaking mechanism was proposed.
Hitachi Research Laboratory, Hitachi Ltd., Japan.
M
M
Ishida
R
R
Haga
N
N
Nishimura
H
H
Matuzaki
R
R
Nakano
D000818:Animals; D000934:Antifoaming Agents; D001709:Biotechnology; D002448:Cell Adhesion; D002452:Cell Count; D002460:Cell Line; D002470:Cell Survival; D003470:Culture Media; D003584:Cytological Techniques / Q000295:instrumentation; D010100:Oxygen; D012833:Siloxanes
D000934:Antifoaming Agents; D003470:Culture Media; D012833:Siloxanes; D010100:Oxygen