Isolation and characterization of human thyroid endothelial cells.
From collagenase digests of human thyroid, endothelial cells were separated from follicular cells by their greater adherence to gelatin-coated plates. Endothelial cells were further purified using fluorescence-activated cell sorting, selecting for cells expressing factor VIII-related antigen. Isolated cells were negative for thyroglobulin and calcitonin when examined by immunostaining. The receptor for the angiopoietins, Tie-2, was expressed by the cells, and expression was increased by agents that elevate cAMP. Nitric oxide synthase (NOS) 3, the endothelial form of NOS, was expressed by the cells and similarly regulated. Cells responded strongly to the mitogen fibroblast growth factor (FGF)-2 in growth assays but only weakly to vascular endothelial growth factor (VEGF). VEGF was, however, able to stimulate nitric oxide release from the cells consistent with their endothelial origin. The FGF receptor (FGFR1) was full length (120 kDa) and immunolocalized to the cytosol and nucleus. Thyrotropin (TSH) did not regulate FGFR1, but its expression was increased by VEGF. Thrombospondin, a product of follicular cells, was a growth inhibitor, but neither TSH nor 3,5,3'-triiodothyronine had direct mitogenic effects. Thyroid follicular cell conditioned medium contained plasminogen activator activity and stimulated the growth of the endothelial cells, but when treated with plasminogen to produce the endothelial-specific inhibitor, angiostatin, growth was inhibited. Human thyroid endothelial cell cultures will be invaluable in determining the cross talk between endothelial and follicular cells during goitrogenesis.
Patel
Vimal A
VA
Division of Medical Sciences, University of Birmingham, United Kingdom.
Logan
Ann
A
Watkinson
John C
JC
Uz-Zaman
Saad
S
Sheppard
Michael C
MC
Ramsden
James D
JD
Eggo
Margaret C
MC
D015153:Blotting, Western; D002116:Calcitonin / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation; D017364:Collagenases; D017077:Culture Media, Conditioned; D000242:Cyclic AMP / Q000378:metabolism; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005434:Flow Cytometry; D006044:Goiter, Nodular; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000032:analysis; D019001:Nitric Oxide Synthase / Q000032:analysis; D052250:Nitric Oxide Synthase Type III; D010958:Plasminogen / Q000494:pharmacology; D010960:Plasminogen Activators / Q000032:analysis; D011518:Proto-Oncogene Proteins; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis; D051496:Receptor, Fibroblast Growth Factor, Type 1; D042787:Receptor, TIE-2; D017468:Receptors, Fibroblast Growth Factor / Q000032:analysis; D013954:Thyroglobulin / Q000032:analysis; D013961:Thyroid Gland / Q000098:blood supply*; D013972:Thyrotropin / Q000494:pharmacology; D014284:Triiodothyronine / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014841:von Willebrand Factor / Q000032:analysis
D017077:Culture Media, Conditioned; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; C105373:MEN1 protein, human; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D017468:Receptors, Fibroblast Growth Factor; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014841:von Willebrand Factor; D014284:Triiodothyronine; D016222:Fibroblast Growth Factor 2; D010958:Plasminogen; D013972:Thyrotropin; D002116:Calcitonin; D013954:Thyroglobulin; D000242:Cyclic AMP; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496348:FGFR1 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D042787:Receptor, TIE-2; D010960:Plasminogen Activators; D017364:Collagenases