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| Composition | Volume | Final concentration |
|---|---|---|
| Serum free medium (StemLine, SIGMA-Aldrich) | - | |
| GM-CSF (100 µg/ml) | 1/5000 dilution | 20 ng/ml |
| IL-4 (100 µg/ml) | 1/5000 dilution | 20 ng/ml |
| TNF-α (50 µg/ml) | 1/20000 dilution | 2.5 ng/ml |
| Ex-Cyte | 1/500 dilution | 2 µl/ml |
Volume of the medium will vary depending on input cell number. In this protocol, we use 1 ml of medium for 105 cells. The complete medium is prepared freshly and all cytokines and supplements are added right before use.
