Supplement	Stock concentration and solvent	Final concentration	Volume/500 mL medium
Calcium chloride for KGM	0.2 M∗ in sterile H2O	0.20 mM	0.5 mL
Calcium chloride for KGM-LC	0.2 M∗ in sterile H2O	0.06 mM	0.15 mL
Epidermal growth factor (EGF)	1 μg/mL in sterile HBS	1 ng/mL	0.5 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
Bovine pituitary extract (BPE)∗∗	100%	0.2%	1 mL
Penicillin/Streptomycin/Fungizone (PSF)	100×	1×	5 mL

∗The calcium chloride 0.2M stock solution is provided with purchase of Medium 154CF, or can be made in-house.

∗∗We recommend centrifuging BPE to remove particulates prior to freezing aliquots to use in supplementing media. While not a required step, this will significantly reduce potential for debris that causes clouding of media (which could otherwise be confused with microbial contamination). Centrifuge for 20 min at 16,000 rpm (∼20,000 x g), 4°C, then sterile filter through a 0.45 μm filter, then finally a 0.2 μm filter, prior to freezing aliquots at -80°C.