Difference between revisions of "Culturing S2 Cells"
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==Materials== | ==Materials== | ||
*'''S2 cells''' (available as frozen stocks from the Drosophila Genomics Resource Center or vendors) | *'''S2 cells''' (available as frozen stocks from the Drosophila Genomics Resource Center or vendors) | ||
| − | *'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) | + | *'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) - stocked downstairs |
| − | *'''Penicillin/streptomycin''' | + | *'''Penicillin/streptomycin''' 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs |
| − | *'''Insect Cell Media'''. Filter together Media (500 mL) and Pen/Strep ( | + | *'''Insect Cell Media'''. Filter together Media (500 mL) and Pen/Strep (10 mL) into a tissue culture bottle |
==Protocol== | ==Protocol== | ||
Revision as of 16:00, 31 July 2009
Materials
- S2 cells (available as frozen stocks from the Drosophila Genomics Resource Center or vendors)
- Sf-900 II SFM medium (Invitrogen, cat. no. 10902) - stocked downstairs
- Penicillin/streptomycin 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs
- Insect Cell Media. Filter together Media (500 mL) and Pen/Strep (10 mL) into a tissue culture bottle
Protocol
- 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.
- To split, gently aspirate the media (some cells will have detached from the plate surface)
- Pipet a gentle stream of media over the surface of the plate (5 mL for normal passage)
- Add 10 mL to fresh 10 cm dishes
- Add 1 mL of detached cells to each new plate
References
see PMID 11752672 and PMID 18388942
