Recipe

Chemically Defined Medium (CDM)

Reagent Amount Final concentration
F-12 Nutrient Mixture (Ham) with GlutaMAX (Invitrogen 31765-035) 250 mL
Iscove’s modified Dulbecco’s medium (IMDM) with GlutaMAX (Invitrogen 31980-030) 250 mL
BSAa fraction V (e.g., Equitech-Bio, Inc., BAC62 or Sigma-Aldrich A1470)  2.5 g  5 mg/mL
Thioglycerol (11.5 m stock; Sigma-Aldrich M7522)  20 µL  450 µm
Insulin (10 mg/mL stock; Roche 11376497001) 350 µL  7 µg/mL
Transferrin (30 mg/mL stock; Roche 10652202001) 250 µL 15 µg/mL
Chemically defined lipid concentrate (100× stock; Invitrogen 11905-031)   5 mL    1×
Store the medium (final 500 mL) at 4°C and use within 1 mo.
aBSA may need to be batch-tested.
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  1. doi:10.1101/pdb.rec095877 Cold Spring Harb Protoc 2017: pdb.rec095877- © 2017 Cold Spring Harbor Laboratory Press

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