Lymphocyte collection medium

Reagents	Final concentration	Amount
RPMI with L-glutamine	n/a	229.6 mL
Heat-inactivated FBS	50%	250 mL
Anti-mycotic and antibiotic solution (AAS)	1%	5 mL
HEPES buffer	1%	5 mL
Non-essential amino acids (NEAA)	1%	5 mL
Sodium pyruvate	1%	5 mL
Beta mercaptoethanol	0.08%	400 μL
Total		500 mL

Prepare the collecting medium under sterile conditions using a biosafety cabinet. After combining all reagents, filter the medium using a 0.22 μm filter (vacuum filtration flask) and store at 4°C until use within three weeks.