Difference between revisions of "Culturing S2 Cells"
From Bridges Lab Protocols
Davebridges (Talk | contribs) m |
Davebridges (Talk | contribs) m (→Materials) |
||
Line 1: | Line 1: | ||
==Materials== | ==Materials== | ||
− | *'''S2 cells''' | + | *'''S2 cells''' see [[Freezing and Thawing S2 Cells]] |
*'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) - stocked downstairs | *'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) - stocked downstairs | ||
*'''Penicillin/streptomycin''' 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs | *'''Penicillin/streptomycin''' 50X mix (Invitrogen, cat. no. 15240062) - stocked downstairs |
Revision as of 16:03, 31 July 2009
Materials
- S2 cells see Freezing and Thawing S2 Cells
- 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