Table 2.
Recommended ranges for growth factors and small molecule inhibitors added to the basal medium at the time of feeding. Aliquoted growth factors are stored at −80°C. Once thawed, the factors are stored at 4°C until expended.
| Component | Stock Concentration | Medium Concentration |
Source/Catalog # |
|---|---|---|---|
| TGFα (T) | 10 μg/ml in PBS/0.1% BSA | 10 ng/ml | Peprotech/100-16A |
| FGF2 (F) | 100 μg/ml in PBS/0.1% BSA | 50-100 ng/ml | Peprotech/100-18B |
| alternatively, FGF9 | 100 μg/ml in PBS/0.1% BSA | 50-100 ng/ml | Peprotech/100-23 |
| LIF (L) | 10 μg/ml | 1-5 ng/ml | EMD Millipore/LIF2010 |
| Y27632 (Y) | 10 mM in DMSO | 10 μM | Tocris/1254 |
| CHIR 99021 (C) | 10 mM in DMSO | 0.5-1.5 μM | Tocris/4423 |
| Bmp7 (B) | 100 μg/ml in 0.004 N HCl with 0.1% BSA |
5-30 ng/ml | RND Systems/ 5666-BP-010 |
| DAPT (D) | 10 mM in DMSO | 0.5-1.0 μM | EMD Millipore/565770 |