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The synthesis of coagulation factors, including procoagulant factor VIII (FVIIIC) and factor VIII related antigen (FVIIIRAG) was studied using the isolated perfused rat liver. The perfusion fluid consisted of rat blood cells suspended in Tyrodes solution containing bovine albumin. Synthetic function was confirmed by the addition of 35S l-methionine to the perfusion medium. Using two dimensional crossed Immunoelectrophoresis and autoradiography progressive incorporation of radioactivity into plasma protein was demonstrated. This was inhibited by cyclohexamide. Coagulation factor assays demonstrated the synthesis of factors II, IX and X and of factors V and VIIIC. Demonstration of synthesis of FVIIIRAG was attempted using an immunoradiometric assay and a cross-reacting rabbit anti-human antiserum. FV and FVIIIRAG synthesis was more clearly demonstrated when the perfusion fluid was depleted of leucocytes and platelets. Addition of Warfarin inhibited the synthesis of factors II, IX and X but not of factors V and VIII. Cyclohexamide completely inhibited synthesis of all coagulation factors but actinomycin acted only after a latent period. No coagulation factor synthesis was demonstrated in the perfusion system without a liver. The results confirmed the functional capacity of the isolated perfused liver for synthesising proteins and the vitamin K dependent factors and demonstrated similar kinetic features for the synthesis of factors V, VIIIC and, possibly, VIIIRAG.

Synthesis of Coagulation Factors and Factor VIII Related Antigen by the Isolated Perfused Rat Liver