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Optimization of the growth conditions of Sf21 insect cells for high-density perfusion culture in stirred-tank bioreactors

1994; Elsevier BV; Volume: 16; Issue: 6 Linguagem: Inglês

10.1016/0141-0229(94)90022-1

1879-0909

Sven Deutschmann, Volker Jäger,

Insect Resistance and Genetics

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 2.5gl−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 × 107 viable cells ml−1 was achieved.