Culturing S2 Cells

• S2 cells (available as frozen stocks from the Drosophila Genomics Resource Center or vendors)
• Sf-900 II SFM medium (Invitrogen, cat. no. 10902)
• Penicillin/streptomycin 100X mix (Invitrogen, cat. no. 15240062)
• Insect Cell Media. Filter together Media (500 mL) and Pen/Strep (5 mL) into a tissue culture bottle

• Typically cells are split every 3-4 days at a ratio of 1:5 and grown at 25C
• After 2-3 days the cells become slightly less adherent.

1. To split, gently aspirate the media (some cells will have detached from the plate surface)
2. Pipet a gentle stream of media over the surface of the plate (5 mL for normal passage)
3. Add 10 mL to fresh 10 cm dishes
4. Add 1 mL of detached cells to each new plate

see PMID 11752672 and PMID 18388942