Isolation and culture of human gastric endothelial cells

Abstract

Background and Aims: Angiogenesis plays an important role in gastric ulcer healing. Considerable heterogeneity exists between endothelial cells from different blood vessels in vitro. Hitherto, it has not been possible to study gastric angiogenesis using relevant endothelial cells. The aim of this study was to isolate and culture human gastric endothelial (HUGE) cells to allow investigation of gastric ulcer angiogenesis. Methods: Gastric mucosa underwent mechanical and enzymatic disruption. Microvessel fragments and endothelial cells were selected using superparamagnetic beads (Dynabeads; Dynal, Wirral, England) coated with a murine monoclonal anti-human platelet- endothelial cell adhesion molecule 1 antibody. Contaminating nonendothelial cells were removed by mechanical weeding and further Dynabead separation of endothelial cells. Results: HuGE cells have been cultured up to passage 19. HuGE cells formed contact-inhibited monolayers on gelatin-coated surfaces and tube-like structures on basement membrane matrices. Indirect immunofluorescence studies confirmed the endothelial nature of these cells. Basic and acidic fibroblast growth factors and vascular endothelial growth factor promoted proliferation (but not epidermal growth factor). HuGE cells synthesized less prostaglandin I2 and E2 compared with human umbilical vein endothelial cells. Conclusions: Pure cultures of HuGE cells can be obtained. Physiologically important differences from large vessel endothelial cells require further investigation.