Drosophila Schneider 2 (S2) cell culture
Lab/Group: Granucci lab (University of Milano-Bicocca)
Related Journal & Article Information
Journal: Nature
Article Title: CD14 regulates the dendritic cell life cycle after LPS exposure through NFAT activation
Introduction
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.
Materials
Reagents
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
Equipment
Procedure
1.Seed S2 cells at a concentration of 1 × 106 cells/ml in 100 × 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 × 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.
Troubleshooting
Critical Steps
Anticipated Results
References
Acknowledgements
Keywords
Drosophila Schneider 2 cells, 2 cells