The Effect of Corticosteroids on Human Choroidal Endothelial Cells: A Model to Study Central Serous Chorioretinopathy
| Autor: | Szymon M. Kielbasa, Onno C. Meijer, Mahmoud Habeeb, Elon H.C. van Dijk, Roula Tsonaka, H.A.B. Peters, Aikaterini Nikolaou, Silvère M. van der Maarel, Camiel J. F. Boon, Arjanneke F van de Merbel, Hetty C M Sips, Robbert G E Notenboom, Eiko K. de Jong, Paul H.A. Quax, Joost Brinks |
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| Přispěvatelé: | Ophthalmology, ANS - Cellular & Molecular Mechanisms |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
CD31 primary endothelial cell isolation medicine.drug_class human choroidal endothelial cells Protein Serine-Threonine Kinases Models Biological Dexamethasone Sensory disorders Donders Center for Medical Neuroscience [Radboudumc 12] Immediate-Early Proteins Tacrolimus Binding Proteins 03 medical and health sciences 0302 clinical medicine Mineralocorticoid receptor Glucocorticoid receptor All institutes and research themes of the Radboud University Medical Center Downregulation and upregulation glucocorticoid receptor Humans Medicine Aldosterone Glucocorticoids Cells Cultured mineralocorticoid receptor Aged Aged 80 and over Dose-Response Relationship Drug Choroid Immunomagnetic Separation business.industry Endothelial Cells Period Circadian Proteins Mifepristone Cadherins Flow Cytometry Tissue Donors Endothelial stem cell Serous fluid 030104 developmental biology Central Serous Chorioretinopathy Gene Expression Regulation Mineralocorticoid 030221 ophthalmology & optometry Cancer research business hormones hormone substitutes and hormone antagonists Transcription Factors medicine.drug |
| Zdroj: | Investigative Ophthalmology and Visual Science, 59, 13, pp. 5682-5692 Investigative Ophthalmology & Visual Science Investigative ophthalmology & visual science, 59(13), 5682-5692. Association for Research in Vision and Ophthalmology Inc. Investigative Ophthalmology and Visual Science, 59, 5682-5692 Investigative Ophthalmology & Visual Science, 59(13), 5682-5692 |
| ISSN: | 0146-0404 5682-5692 |
| Popis: | Purpose To isolate, culture, and characterize primary human choroidal endothelial cells, and to assess their responsiveness to corticosteroids, in order to enable knowledge gain on the pathogenesis of central serous chorioretinopathy. Methods Choroidal endothelial cells were isolated from cadaveric human donors. Magnetic-activated cell sorting with anti-human CD31 was performed for choroidal endothelial cell isolation. Primary cultures of purified choroidal endothelial cells were treated with several regimens of corticosteroids and analyzed for effects on primary corticosteroid responsive genes. Results Isolated choroidal endothelial cell cultures had a cobblestone appearance in monolayer cultures and stained positive for vascular endothelial cadherin. Moreover, on a 3D-Matrigel matrix, these cells formed capillary-like structures, characteristic of in vitro endothelial cells. Primary cultures of purified choroidal endothelial cells treated with several regimens of corticosteroids demonstrated significant transcriptional upregulation of primary corticosteroid responsive genes (FKBP5, PER1, GILZ, and SGK1). Further pharmacologic analysis using specific agonists (dexamethasone, aldosterone) and antagonists (mifepristone, spironolactone) for either the glucocorticoid receptor or the mineralocorticoid receptor showed that this response was exclusively mediated by the glucocorticoid receptor in our model. Conclusions With this optimized choroidal endothelial cell isolation and culturing protocol, we have established an in vitro model that appears very suitable for research on both central serous chorioretinopathy and other diseases in which corticosteroids and choroidal endothelial cells are involved. Our model proves to be suitable for studying effects mediated through the glucocorticoid receptor. The role of mineralocorticoid receptor-mediated effects needs further research, both in vivo and in cell model development. |
| Databáze: | OpenAIRE |
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