Culturing S2 Cells: Difference between revisions
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*'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) | *'''Sf-900 II SFM medium''' (Invitrogen, cat. no. 10902) | ||
*'''Penicillin/streptomycin''' 100X mix (Invitrogen, cat. no. 15240062) | *'''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 | |||
==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== | ==References== | ||
see PMID 11752672 and PMID 18388942 | see PMID 11752672 and PMID 18388942 | ||
[[Category: Cell Culture]] | [[Category: Cell Culture]] | ||
[[Category: Drosophila]] | [[Category: Drosophila]] | ||
Revision as of 15:43, 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)
- 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
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