Abstract
We developed a method for long-term culture of microvascular endothelial cells derived from mongolial gerbil brain, and investigated their biological properties in vitro. Microvessels were isolated from mongolial gerbil brain by combination of the enzymatic treatment, filtration and centrifugation, and seeded onto a gelatin coated dish. A morphologically homogeneous cell plaque was removed two to three weeks after the seeding, and the obtained cells were subcultured. The cultured cells grew as monolayers of flat polygonal cells, and sometimes formed cellular cords. The cultured cells were carried more than 20 passages without morphological change, and have retained an endothelial specific marker, Factor VIII-related antigen even in late passages. Histochemical activity of alkaline phosphatase was present in almost all cells as an intense diffuse and granular staining in primary culture. The activity slowly disappeared as the cells proliferate, and almost completely disappeared after fifth passage. Activity of γ-glutamyl transpeptidase was demonstrated only in the cells at the center of the cell plaque sorrounded by non-reactive cells in primary culture. There was no activity in the cells after the second passage. The membrane specific enzyme for cerebral microvessel seemed to disappear when the cells migrate or proliferate from the microvessel isolates.
