KSOM-AA
Glucose, 0.04 g/l
Penicillin G, 0.06 g/l
Streptomycin, 0.05 g/l
Sodium lactate, 1.12 g/l (or 1.87 g of 60% syrup per liter)
NaHCO3, 2.1 g/l
Phenol red, 0.1 ml of 0.5% or 1% solution per liter
Sodium pyruvate, 0.02 g/l
L-glutamine, 0.146 g/l
To make KSOM-AA: Supplement KSOM with amino acids. Stock solutions of minimal essential medium (MEM) essential acids (EAA) and nonessential acids (NEAA) can be used (e.g., Invitrogen). Use NEAA and EAA at half of Eagle's concentations, i.e., add 0.5 ml NEAA and 1 ml EAA per 100 ml of KSOM.
Osmolarity should be 256 mOsm. Filter sterilize through a 0.2 μm Millipore filter. Aliquot in polypropylene tubes and store at 4°C for 1-2 weeks.
Note:
Phenol red is optional; it may be embryotoxic.
Note:
2x KSOM can be prepared without BSA and CaCl2 and stored at -70°C for up to 3 months. 100 mg/ml BSA and 0.171 M CaCl2 stocks (100x) are frozen separately and added at the time of medium preparation.
