Hormone-Containing Medium

Recipe

Reagent	Final concentration
Leibovitz's L-15 medium	60%
Charcoal-treated serum (CTS) <R>	10%
3,3′,5-triiodo-l-thyronine (T3) (Sigma-Aldrich T6397; 404 µg/mL in 0.01 m NaOH)	10 nm
Insulin bovine (Sigma-Aldrich I664; 5 mg/mL in 0.005 n HCl)	5 µg/mL
Hydrocortisone (Sigma-Aldrich H0888; 0.5 mg/mL in distilled water)	0.5 µg/mL
Penicillin (10,000 U/mL)	100 U/mL
Streptomycin (10,000 µg/mL)	100 µg/mL
Just before use, combine reagents to the desired volume in sterilized distilled water, and filter sterilize through a 0.22-µm filter.