M-199 incubation medium

Ficoll 400 (Sigma)

Medium 199, modified with Hanks’, L-glutamine (US Biological M2852)

• 1. Add 2 g of Ficoll 400 to 100 mL of Medium 199. Mix with a magnetic stir bar until the Ficoll dissolves.

• 2. If necessary, adjust the pH to 7.2-7.5 with NaOH.

• 3. Filter-sterilize with a Nalgene 115-mL sterile filter unit type S (0.20-μm) or a 10-mL syringe attached to a 0.2-μm syringe filter.

• 4. Store refrigerated in labeled 15-mL vials.

  After prolonged storage, refilter the medium through a 0.2-μm syringe filter.