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Isolation and characterization of human thyroid endothelial cells
Vimal A Patel1, Ann Logan, John C Watkinson
1Division of Medical Sciences, University of Birmingham, United Kingdom.
American Journal of Physiology. Endocrinology and Metabolism
|October 22, 2002
Summary
Researchers isolated human thyroid endothelial cells, finding they respond to fibroblast growth factor (FGF)-2 and release nitric oxide. These cells are crucial for understanding thyroid goitrogenesis and cell communication.
Area of Science:
- Endocrinology
- Cell Biology
- Vascular Biology
Background:
- Thyroid endothelial cells play a role in thyroid function and disease.
- Understanding the specific characteristics and functions of these cells is essential.
Purpose of the Study:
- To isolate and characterize human thyroid endothelial cells.
- To investigate their response to growth factors and signaling molecules.
- To establish a model for studying endothelial-follicular cell interactions in goitrogenesis.
Main Methods:
- Isolation of endothelial cells from human thyroid tissue using differential adherence to gelatin-coated plates.
- Purification via fluorescence-activated cell sorting (FACS) for factor VIII-related antigen.
- Immunostaining to confirm absence of follicular cell markers (thyroglobulin, calcitonin).
- Assessment of receptor expression (Tie-2, FGFR1) and protein localization.
- Cell growth assays in response to fibroblast growth factor (FGF)-2 and vascular endothelial growth factor (VEGF).
- Nitric oxide synthase (NOS) activity and nitric oxide release measurements.
- Analysis of thyroid follicular cell conditioned medium effects.
Main Results:
- Isolated cells expressed Tie-2 and nitric oxide synthase (NOS) 3, with Tie-2 expression upregulated by cAMP-elevating agents.
- Cells exhibited strong proliferation in response to FGF-2 and weak response to VEGF, but VEGF stimulated nitric oxide release.
- Fibroblast growth factor receptor (FGFR1) was detected in the cytosol and nucleus.
- Thyroid follicular cell conditioned medium stimulated endothelial cell growth, an effect inhibited by angiostatin derived from plasminogen.
Conclusions:
- Successfully isolated and characterized functional human thyroid endothelial cells.
- Demonstrated specific growth factor responses and nitric oxide production capabilities.
- Established a foundation for investigating the cross-talk between thyroid endothelial and follicular cells in the context of goitrogenesis.