Protocol for Drosophila Schneider 2 (S2) cell culture used in our Nature paper.
S2 cells (macrophage-like lineage) grow at room temperature without CO2, as a loose, semi-adherent monolayer in tissue culture flasks and plates.
Method Article
Drosophila Schneider 2 (S2) cell culture
https://doi.org/10.1038/nprot.2009.135
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Protocol for Drosophila Schneider 2 (S2) cell culture used in our Nature paper.
S2 cells (macrophage-like lineage) grow at room temperature without CO2, as a loose, semi-adherent monolayer in tissue culture flasks and plates.
Complete Schneider’s Drosophila medium recipe:
HI FBS (EuroClone) - 10%
Penicillin/Streptomycin (EuroClone) - 50 U/ml
Conditioned complete Schneider’s medium - 25%
Schneider’s Drosophila medium (Gibco) - to volume
1.Seed S2 cells at a concentration of 1 x 106 cells/ml in 100 x 20 mm tissue culture plates in 10 ml of conditioned medium (supplemented with 25% of the medium in which cells have been formerly grown).
2.Incubate in a 28°C incubator (no CO2 requirements) and split cells at 1:2 to 1:5 dilution when cell density is between 6 to 20 x 106 cells/ml.
3.If planning to perform LPS or PGN stimulation on S2 cells, pretreat cells with 1 microM 20-hydroxy-ecdysone for 24 hours before stimulation.
This protocol has been posted on Protocol Exchange, an open repository of community-contributed protocols sponsored by Nature Portfolio. These protocols are posted directly on the Protocol Exchange by authors and are made freely available to the scientific community for use and comment.
posted
You are reading this latest protocol version