In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology
A co-culture assay with human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblasts (NHDFs) was used to study whether selected angiogenesis inhibitors were able to inhibit differentiation and network formation of HUVECs in vitro. The effect of the inhibitors was determined by...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-04-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/8/4277 |
| _version_ | 1797445938200969216 |
|---|---|
| author | Arlinda Ljoki Tanzila Aslam Tina Friis Ragnhild G. Ohm Gunnar Houen |
| author_facet | Arlinda Ljoki Tanzila Aslam Tina Friis Ragnhild G. Ohm Gunnar Houen |
| author_sort | Arlinda Ljoki |
| collection | DOAJ |
| description | A co-culture assay with human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblasts (NHDFs) was used to study whether selected angiogenesis inhibitors were able to inhibit differentiation and network formation of HUVECs in vitro. The effect of the inhibitors was determined by the morphology and the calculated percentage area covered by HUVECs. Neutralizing VEGF with avastin and polyclonal goat anti-VEGF antibody and inhibiting VEGFR2 with sorafenib and vatalanib resulted in the formation of HUVEC clusters of variable sizes as a result of inhibited EC differentiation. Furthermore, numerous inhibitors of the VEGF signaling pathways were tested for their effect on the growth and differentiation of HUVECs. The effects of these inhibitors did not reveal a cluster morphology, either individually or when combined to block VEGFR2 downstream pathways. Only the addition of <i>N</i>-methyl-<i>p</i>-bromolevamisole revealed a similar morphology as when targeting VEGF and VEGFR2, meaning it may have an inhibitory influence directly on VEGFR signaling. Additionally, several nuclear receptor ligands and miscellaneous compounds that might affect EC growth and differentiation were tested, but only dexamethasone gave rise to cluster formation similarly to VEGF-neutralizing compounds. These results point to a link between angiogenesis, HUVEC differentiation and glucocorticoid receptor activation. |
| first_indexed | 2024-03-09T13:33:00Z |
| format | Article |
| id | doaj.art-4510bff6ff014d8281cb6dccb01fde4d |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:33:00Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-4510bff6ff014d8281cb6dccb01fde4d2023-11-30T21:15:25ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-04-01238427710.3390/ijms23084277In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and MorphologyArlinda Ljoki0Tanzila Aslam1Tina Friis2Ragnhild G. Ohm3Gunnar Houen4Department of Autoimmunology, Statens Serum Institut, DK-2300 Copenhagen, DenmarkDepartment of Autoimmunology, Statens Serum Institut, DK-2300 Copenhagen, DenmarkDepartment of Autoimmunology, Statens Serum Institut, DK-2300 Copenhagen, DenmarkDepartment of Drug Design and Pharmacology, University of Copenhagen, DK-2200 Copenhagen, DenmarkDepartment of Autoimmunology, Statens Serum Institut, DK-2300 Copenhagen, DenmarkA co-culture assay with human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblasts (NHDFs) was used to study whether selected angiogenesis inhibitors were able to inhibit differentiation and network formation of HUVECs in vitro. The effect of the inhibitors was determined by the morphology and the calculated percentage area covered by HUVECs. Neutralizing VEGF with avastin and polyclonal goat anti-VEGF antibody and inhibiting VEGFR2 with sorafenib and vatalanib resulted in the formation of HUVEC clusters of variable sizes as a result of inhibited EC differentiation. Furthermore, numerous inhibitors of the VEGF signaling pathways were tested for their effect on the growth and differentiation of HUVECs. The effects of these inhibitors did not reveal a cluster morphology, either individually or when combined to block VEGFR2 downstream pathways. Only the addition of <i>N</i>-methyl-<i>p</i>-bromolevamisole revealed a similar morphology as when targeting VEGF and VEGFR2, meaning it may have an inhibitory influence directly on VEGFR signaling. Additionally, several nuclear receptor ligands and miscellaneous compounds that might affect EC growth and differentiation were tested, but only dexamethasone gave rise to cluster formation similarly to VEGF-neutralizing compounds. These results point to a link between angiogenesis, HUVEC differentiation and glucocorticoid receptor activation.https://www.mdpi.com/1422-0067/23/8/4277angiogenesisco-cultureHUVECinhibitormorphologyNHDF |
| spellingShingle | Arlinda Ljoki Tanzila Aslam Tina Friis Ragnhild G. Ohm Gunnar Houen In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology International Journal of Molecular Sciences angiogenesis co-culture HUVEC inhibitor morphology NHDF |
| title | In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology |
| title_full | In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology |
| title_fullStr | In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology |
| title_full_unstemmed | In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology |
| title_short | In Vitro Angiogenesis Inhibition and Endothelial Cell Growth and Morphology |
| title_sort | in vitro angiogenesis inhibition and endothelial cell growth and morphology |
| topic | angiogenesis co-culture HUVEC inhibitor morphology NHDF |
| url | https://www.mdpi.com/1422-0067/23/8/4277 |
| work_keys_str_mv | AT arlindaljoki invitroangiogenesisinhibitionandendothelialcellgrowthandmorphology AT tanzilaaslam invitroangiogenesisinhibitionandendothelialcellgrowthandmorphology AT tinafriis invitroangiogenesisinhibitionandendothelialcellgrowthandmorphology AT ragnhildgohm invitroangiogenesisinhibitionandendothelialcellgrowthandmorphology AT gunnarhouen invitroangiogenesisinhibitionandendothelialcellgrowthandmorphology |