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RPMI1640, 52.5%; FBS, 40%; DMSO, 7.5%

Dulbecco's modified Eagle's Medium, with 4.5g/L glucose, 85%; heat inactivated fetal bovine serum (FBS), 15%

Dulbecco's modified Eagle's medium, 90%; heat inactivated fetal bovine serum (FBS) 10%

F9 cells can be stimulated to differentiate into parietal endoderm in the presence of retinoic acid and dibutyryl cyclic AMP (cAMP); differentiating cells synthesize plasminogen activator, laminin and type IV collagen; cAMP is active only on cells that have been treated with retinoic acid; tested and found negative for ectromelia virus (mousepox); culture vessels must be coated with 0.1% gelatin prior to use. To do so, cover the surface to of the vessel with 0.1% gelatin (Difco) in sterile distilled water for 2 hours at 4C, then wash three times with sterile distilled water. Treated flasks and dishes can be stored at room temperature