CEPT Cocktail

1. Prepare stock solutions for each compound
a. Prepare 0.5 mM of Chroman 1 stock solution (10,000×): reconstitute 5mg of Chroman 1 in 22.91 mL of DMSO. Vortex solution until the Chroman 1 is completely dissolved.
The working concentration of Chroman 1 is 50 nM.
b. Prepare 50 mM of Emricasan stock solution (10,000×): reconstitute 10mg of Emricasan in 0.3512 mL of DMSO. Vortex solution until the Emricasan is completely dissolved.
The working concentration of Emricasan is 5 μM.
c. Prepare 7 mM of Trans-ISRIB stock solution (10,000×): reconstitute 10mg of Trans-ISRIB in 3.165 mL of DMSO. Gently warm the solution between 45 °C and 60 °C , and vortex solution until the Trans-ISRIB is completely dissolved.
The working concentration of Trans-ISRIB is 0.7 μM.

2. Supplement an aliquot of complete hPSC culture medium with CET small molecules 1:10,000 just before use. For example, add 1 μL of 10,000× stock solution of each small molecule per 10 mL of medium.

3. Add 10 μL of 1,000× Polyamine Solution per 10 mL of medium.

4. Filter the prepared medium using a 0.22 μm filter prior to use.