4. Neubauer improved counting chamber (e.g., Marienfeld,
Ko ¨nigshofen, Germany).
5. Piston pipettes (e.g., Research Plus, Eppendorf, Hamburg,
Germany).
6. Pipette tips (e.g., Sarstedt, Nu ¨mbrecht, Germany).
2.4.3 Materials for Cell
Passaging
1. 100 mL baffled shake flask (e.g., Glasger€ atebau Ochs, Bovenden, Germany).
2. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
3. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
4. Sterile single-use pipette tips (e.g., serological pipettes, Sarstedt, Nu ¨mbrecht, Germany).
5. Incubator (e.g., B-line, Binder, Tuttlingen, Germany) with
orbital shaker (e.g., Celltron, Infors HT, Basel, Switzerland).
2.4.4 Materials
for Freezing the Cells
1. Spodoptera frugiperda cell suspension in exponential growth
phase with a cell density of 2–6 Â 10
6 cells/mL and a viability
>95%.
2. Sterile 1.5 mL Cryo-Vials (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
3. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
4. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
5. Sterile serological pipettes (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
6. 15-mL centrifuge tubes (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
7. Aspiration system (e.g., Vacusafe, Integra, Biebertal,
Germany).
8. Freezing container (e.g., Mr. Frosty, Thermo Fisher Scientific,
Waltham, MA, USA).
9. À80
C freezer.
10. À140
C liquid nitrogen freezing tank.
11. Dimethyl sulfoxide (DMSO).
2.4.5 Materials
for Cultivation of Insect
Cells in a Stirred-Tank
Bioreactor
1. Bioreactor with 1 L working volume (e.g., 2L-Labfors, Infors
HT, Basel, Switzerland).
2. Bioreactor control unit (e.g., Labfors 5 cell, Infors HT, Basel,
Switzerland).
Antigen Production in Insect Cells
101