AI Article Synopsis
- - Endothelial cell tube formation assay is a reliable in vitro method to study angiogenesis, where endothelial cells create a tube network on a basement membrane extract when exposed to specific angiogenic factors.
- - The tube formation occurs rapidly and remains stable for 18-24 hours before cells start to die and the structure disintegrates, with the network being observable through microscopy techniques.
- - The assay offers advantages such as easy setup, the ability to test multiple factors at once, quick results, and quantifiability, but requires careful selection of materials and optimized conditions for consistent results.
Article Abstract
Endothelial cell tube formation assay is one of the most widely used and reliable methods for studying in vitro angiogenesis. Endothelial cells plated over a basement membrane extract and subjected to angiogenic factors in conditioned medium, form a rapid and quantifiable tube network within hours. Tube formation is sustained for 18-24 h, after which time apoptosis occurs and tube networks disintegrate. The tube network can be imaged using a phase contrast microscope, or upon Calcein-AM treatment, a fluorescence/confocal microscope. This assay has several advantages, namely: ease of set up, the ability to test numerous angiogenic/anti-angiogenic factors simultaneously, quick network formation, ability to view live or fixed tube networks, and quantifiability. To ensure successful results and limit variability, proper selection of basement membrane extracts and endothelial cells is necessary, and conditions must be optimized. In summary, this assay is a useful method for screening potential angiogenic/anti-angiogenic factors as well as identifying critical mechanisms and signaling pathways underlying angiogenic-related pathologies.
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| http://dx.doi.org/10.1007/978-1-0716-2217-9_12 | DOI Listing |
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