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| - Isolation, culture, and characterization of endothelial cells derived from the post-capillary venules of sheep mesenteric lymph nodes, Peyer's patches, and associated small bowel
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| - Lymphocyte recirculatlon is mediated principally by specialized endothellal cells which line the post-capillary venules of lymph nodes and other secondary lymphoid tissues. The ontogeny and physiology of this process have been characterized in sheep In considerable detail. To further enhance the analytical potential of this experimental system we have isolated endothelial cells from the post-capillary venules of ovine mesenteric lymph nodes by perfusion with small (37-74 μm diameter), sulfonated microcarrier beads. Cells isolated in this manner have been maintained in vitro for >12 months through >30 passages. The endothellal nature of these cells has been conclusively established on the basis of morphologic, metabolic, and immunologic criteria. Virtualty all (>99%) cells in primary and passaged cultures metabolized Dil-AC-LDL, a known marker for endothellal cells. Furthermore, nearly all (>95%) cells expressed cell-surface von Willebrand factor and antlthrombln III, which are known endothellal antigens. All cells expressed major histocompatibility class I antigens but no cells expressed class II antigens. In vitro lymphocyte-binding studies revealed that these cells bound lymphocytes in a dose-dependent fashion. The microcarrier perfusion technique was also used to isolate endothellal cells from the post-capillary venules of lieal Peyer's patches and associated small bowel in sheep. The majority (70%) of cells isolated in this manner resembled the cells isolated from mesenterlc lymph nodes both morphologically and metabolically.
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