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| Composition | Volume | Final concentration |
|---|---|---|
| Differentiation (hPSC/OP9 coculture) medium | - | |
| GM-CSF (100 µg/ml) | 1/2000 dilution | 50 ng/ml |
| 1α, 25-Dihydroxyvitamin D3 (1 mM) | 1/5000 dilution | 200 nM |
| RANKL (10 µg/ml) | 1/1000 dilution | 10 ng/ml |
Volume of the medium will vary depending on input cell number. In this protocol, we use 1ml of medium for 5×104 cells. The complete medium is prepared freshly and all cytokines and supplements are added right before use.
