ABSTRACT
In this chapter the author describes state-of-the-art approaches to the culture of vascular cells from the lung that have been successfully applied in his and other investigators' laboratories. Human, bovine, and porcine tissues are the most widely used sources of lung endothelial cells. In author's laboratory, bovine cells are used because the tissue can be obtained easily. At most cattle abattoirs, blood vessels from adults or fetuses can be obtained for a nominal fee. Bovine pulmonary endothelial cells are maintained in medium 199 with Earle's salts supplemented as described by Lewis. To prepare endothelial clones, one must use "conditioned" medium to promote cell division in low-density cultures. Conditioned medium is prepared by removing medium from a culture of smooth muscle cells and replacing it with serumless medium. Endothelial cells in vitro show a distinctly squamous epithelial morphology. When freshly isolated endothelial cells are first placed in culture, they initially are associated with one another in the form of small clumps.
