Complete tissue culture medium (CTM)

Reagents	Final concentration	Amount
RPMI with L-glutamine	n/a	500 mL
Heat-inactivated FBS	8.7%	50 mL
Anti-mycotic and antibiotic solution (AAS)	1%	6 mL
HEPES buffer	1%	6 mL
Non-essential amino acids (NEAA)	1%	6 mL
Sodium pyruvate	1%	6 mL
Beta mercaptoethanol	0.08%	450 μL
Total		574.45 mL

Prepare CTM under sterile conditions in a biosafety cabinet. After mixing all reagents, filter the CTM using a 0.22 μm filter (vacuum filtration flask) and store at 4°C until use for about three weeks.