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Sf9 insect cells were adapted prior to growth in suspension or monolayer to serum-free EX-CELL 420 insect cell culture medium (Sigma) at 27 °C. Suspension culture cells were usually seeded at 0.5–1 × 106 cells/mL in a total volume of 25 mL in an upright standing 75 cm2 disposable T-flask. Cells were exponentially grown, incubated in a controlled shaker at 27 °C and 100 rpm. Cell density was counted daily. When the density reached 4 × 106 cells/mL cells were split.
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