Properties of cultured endothelium from adult human vessels.

Abstract

Endothelium was isolated from samples of aorta and vena cava obtained from cadaver donors at the time kidneys were harvested for transplantation. Digestion with collagenase and gentle swabbing were used to free the cells from the intimal surface. Low density seeding permitted isolation of individual colonies with typical endothelial morphology. Modified Medium 199 supplemented with 10%-20% human plasma-derived serum and an extract from the bovine hypothalamus (500 micrograms/ml) enabled subcultured colonies to grow to confluency when culture surfaces were coated with fibronectin (1 micrograms/cm2). The presence of Factor VIII antigen was demonstrated using an indirect immunofluorescence technique. A monoclonal antibody to cultured umbilical vein endothelium, specific for endothelium, reacted with the subcultured cells from the aorta and vena cava. Type IV procollagen, fibronectin, and thrombospondin were identified as labeled proteins secreted by cultures of adult endothelium that had been incubated with 3H-proline and 3H-glycine. When the cultured endothelium was used in a sodium-m-periodate stimulated T lymphocyte mitogenic culture system, the endothelium exhibited accessory cell function. Prostacyclin production stimulated by incubation with arachidonic acid and PGH2 was variable from vessel to vessel. However, average values were lower than normally seen with cultured primary umbilical vein endothelium.