Supplement	Stock concentration and solvent	Final concentration	Volume/500 mL medium
Bovine pituitary extract (BPE)∗∗	100%	0.4%	2 mL
Hydrocortisone	0.5 mg/mL in 100% ethanol	0.5 μg/mL	0.5 mL
Human insulin	5 mg/mL in sterile 10 mM HCl	5 μg/mL	0.5 mL
Triiodothyronine	20 nM in sterile HBS	20 pM	0.5 mL
α-Melanocyte stimulating hormone (αMSH)	1 μM in sterile HBS	1 nM	0.5 mL
Endothelin 1	0.5 μM in sterile HBS	0.5 nM	0.5 mL
Basic fibroblast growth factor (bFGF)	1 μg/mL in 0.1% Bovine Serum Albumin (BSA) in Tris buffer pH 7.6	1 ng/mL	0.5 mL
Penicillin/Streptomycin/Fungizone (PSF)	100×	1×	5 mL

∗∗See previous table for recommended centrifugation of BPE.