DMEM-Sato Base Growth Medium (with NB)

• 1. Combine the following:

  Reagent	Amount (for 20 mL)	Final
  Neurobasal medium (Gibco 21103-049)	9.5 mL
  DMEM (Gibco 11960-044)	9.5 mL
  Insulin stock (0.5 mg/mL)	200 µL	5 µg/mL
  Sodium pyruvate (100 mm; Gibco 11360-070)	200 µL	110 µg/mL
  Penicillin-streptomycin (100×; Gibco 15140-122)	200 µL	100 U/mL (penicillin)
  		100 µg/mL (streptomycin)
  SATO supplement (100×)	200 µL	1×
  Thyroxine (T3) stock (4 µg/mL)	200 µL	40 ng/mL
  l-glutamine (200 mm; Gibco 25030-081)	200 µL	292 µg/mL
  NS21 supplement (50×) (R&D Systems AR008; see Chen et al. 2008)	400 µL	1×
  NAC stock (5 mg/mL)	20 µL	5 µg/mL

• 2. Filter through a rinsed 0.22-µm filter to sterilize.

• © 2013 Cold Spring Harbor Laboratory Press