Application of CelliGen 310 Bioreactor to Sf9 Cells

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Application of CelliGen® 310 Bioreactor to Sf9 Cells
Richard Mirro
New Brunswick, Enfield, CT USA
Introduction
The Sf9 insect cell-baculovirus system is widely used for the expression of various recombinant proteins. The cells are non-adherent-dependent cells suitable for high-density, large-scale suspension culture. The use of bioreactors to culture Sf9 cells can well control the cell growth environment and greatly increase cell density and protein expression. At present, the suspension culture method of Sf9 cells in the reactor is mainly in the form of Fed-batch. The method is to supplement the nutrients with the increase of cell density and nutrient consumption, but no effluent of culture solution. Device. There is also a continuous method in which the in-tank culture solution is discharged at the same rate according to the continuous addition of fresh medium. This article mainly introduces the cultivation of Sf9 cells in a continuous culture mode.
Method and apparatus
Bioreactor: New Brunswick CelliGen 310, Spin Filter Impeller with Rotary Filter, Working Volume: 3.5 L
Cell: Sf9
Medium: Sf-900 serum-free medium, GIBCO cell seeding density: 4.0-5.0×105/ml
Perfusion rate: 0-0.5 working volume (according to cell density)
result

Figure 1: Schematic diagram of cell density and viable cell rate on day 8

Figure 2: Mixer paddle with rotary filter

Sf9 reached a maximum density of 2.5 × 107 cells/ml after 8 days of growth in the bioreactor, while the viable cell rate remained above 90% (Fig. 1). The maximum density of 6-12×106 cells/ml achieved by batch culture is increased by 2-4 fold, which provides a basis for high-density, large-scale continuous culture of Sf9 cells in bioreactors.
discuss
The cell-carrying mesh (10 μm) with a rotating filter screw agitator (Fig. 2) can retain Sf9 cells in the reactor and continue to grow. The cell-free supernatant is perfused out of the reactor while replenishing fresh culture. base. On the one hand, this method provides nutrients required by high-density cells; on the other hand, it dilutes the harmful products produced by cell metabolism, reduces the negative effects on cells, and finally achieves high-density culture of Sf9 cells.

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