Difference between revisions of "Culturing S2 Cells"

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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)
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*'''Insect Cell Media'''.  Filter together Media (500 mL) and Pen/Strep (5 mL) into a tissue culture bottle
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 +
==Protocol==
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*Typically cells are split every 3-4 days at a ratio of 1:5 and grown at 25C
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*After 2-3 days the cells become slightly less adherent.
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#To split, gently aspirate the media (some cells will have detached from the plate surface)
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#Pipet a gentle stream of media over the surface of the plate (5 mL for normal passage)
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#Add 10 mL to fresh 10 cm dishes
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#Add 1 mL of detached cells to each new plate
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==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.
  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

References

see PMID 11752672 and PMID 18388942