Synthesis and Release of Factor VIII by Cultured Human Endothelial Cells

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Abstract
Summary. Endothelial cells (ECs) derived from human umbilical veins were cultured in order to study the physiological control of factor VIII synthesis and release. The culture media were studied from multiple replicate cultures at confluence. Factor VIII related antigen (VIIIR:Ag) and factor VIII coagulant antigen (VIII:CAg) were measured by sensitive immunoradiometric assays. De novo synthesis of factor VIII related protein (VIII:R) was quantitated by incorporation of labelled amino acids into specific protein subunits. The following agents were added to the culture medium in a range of concentrations from physiological to pharmacological: adrenaline, 5 hydroxytryptamine, 2,3-DPG, cyclic AMP, thyroxine, hydrocortisone, and human growth hormone. None of them had any effect at any concentration on the rate of accumulation of VIIIR:Ag in the culture medium. Addition of exogenous factor VIII had no effect on do novo synthesis of VIII:R. VIII:CAg was found to be stable under the conditions of culture but none was released from the ECs. Long-term monocyte cultures also failed to release VIII:CAg. It appears that VIII:R is a constitutive gene product of umbilical vein endothelial cells and that VIII:CAg is not made by these cells.