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.
Lymphocyte recirculation is mediated principally by specialized endothelial 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 microns diameter), sulfonated microcarrier beads. Cells isolated in this manner have been maintained in vitro for greater than 12 months through greater than 30 passages. The endothelial nature of these cells has been conclusively established on the basis of morphologic, metabolic, and immunologic criteria. Virtually all (greater than 99%) cells in primary and passaged cultures metabolized Dil-AC-LDL, a known marker for endothelial cells. Furthermore, nearly all (greater than 95%) cells expressed cell-surface von Willebrand factor and antithrombin III, which are known endothelial 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 endothelial cells from the post-capillary venules of ileal Peyer's patches and associated small bowel in sheep. The majority (70%) of cells isolated in this manner resembled the cells isolated from mesenteric lymph nodes both morphologically and metabolically.
Abernethy
N J
NJ
Department of Pathology, University of Toronto, Ontario, Canada.
Hay
J B
JB
D000818:Animals; D002465:Cell Movement; D002469:Cell Separation; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D007421:Intestine, Small / Q000098:blood supply; D008077:Lipoproteins, LDL / Q000378:metabolism; D008198:Lymph Nodes / Q000098:blood supply; D008214:Lymphocytes / Q000166:cytology / Q000502:physiology; D010581:Peyer's Patches / Q000098:blood supply; D012756:Sheep; D014841:von Willebrand Factor / Q000378:metabolism
D008077:Lipoproteins, LDL; C032629:acetyl-LDL; D014841:von Willebrand Factor