Optimization of the growth conditions of Sf21 insect cells for high-density perfusion culture in stirred-tank bioreactors.
Spodoptera frugiperda insect cells (IPLB-Sf21-AE) (Sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. In the present study the culture conditions of these insect cells were studied to establish high-density suspension cultures with prolonged exponential growth phases. The Sf21 cells were grown in 125-ml spinner flasks using five different culture media supplemented with 5% fetal calf serum and four protein-free or low-protein culture media. The best results were achieved in EX-CELL 401 (protein-free media) and in IPL-41 modified with 2.5 g l-1 tryptose phosphate broth (serum-supplemented media), respectively. The latter was used for further batch and continuous cultivation of Sf21 cells in a perfused 1.4-l stirred-tank bioreactor with special attention to the oxygen requirement of these cells. Optimal growth was found at an oxygen concentration of 70% air saturation, resulting in a prolonged exponential growth phase that could be maintained for more than 16 days. A maximum cell density of 5.5 x 10(7) viable cells ml-1 was achieved.
Deutschmann
S M
SM
Gesellschaft für Biotechnologische Forschung mbH, Arbeitsgruppe Zellkulturtechnik, Braunschweig, Germany.
Jäger
V
V
D000818:Animals; D016367:Baculoviridae; D001709:Biotechnology / Q000379:methods*; D002455:Cell Division; D002460:Cell Line; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D005822:Genetic Vectors; D007700:Kinetics; D009036:Moths; D010100:Oxygen / Q000494:pharmacology; D010477:Perfusion; D013997:Time Factors
D003470:Culture Media; D010100:Oxygen