Weihua Song1, Chee Wai Fhu1, Koon Hwee Ang2, Cheng Hao Liu1, Nurul Azizah Binte Johari1, Chin Shiuan Lio1, Sabu Abraham3, Wan Jin Hong2, Stephen E Moss4, John Greenwood4, Xiaomeng Wang1,2,4
1 Lee Kong Chian School of Medicine, Nanyang Technological University, 50 Nanyang Drive, Research Techno plaza, Level 4, X-Frontiers Block, Singapore 637664.
2 Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Proteos, Singapore 138673.
3 Institute of Cardiovascular Sciences, 3.20, Core Technology Facility (3rd floor) 46 Grafton Street, Manchester, UK, M13 9NT.
4 Department of Cell Biology, Institute of Ophthalmology, University College London, 11-43 Bath Street, London, UK, EC1V 9EL.
Published online in Nature Protocols on 3 September 2015.
The mouse fetal metatarsal provides a unique tool for studying angiogenesis. In comparison with other commonly used in vitro or ex vivo angiogenesis assays, vessel outgrowth from mouse fetal metatarsals is more representative of sprouting angiogensis in vivo. It allows the analysis of blood vessel growth, and the mechanisms underpinning this process, in a multicellular microenvironment that drives the formation of a robust and complex vascular network in the absence of exogenous growth factors. By labeling different constituents of the vascular structure, it is possible to perform 3D rendering of the spatial interplay between different cellular components and to carry out quantitative analysis of vessel outgrowth. High-resolution imaging permits the visualization of fine structural and cellular details. As the assay involves the use of fetal tissues, it is possible to follow new blood vessel formation in genetically modified mice that are perinatally lethal. The entire process takes 9-13 d. A detailed description of how to set up and perform the assay is described here.
This paper was featured as a cover story in Nature Protocols. Please click here.
Figure legend: The cover image is reproduced from a mixed oil and acrylic on canvas painting by Jestin George. The image depicts the artist's interpretation of the immunostained microvascular network generated during the metatarsal angiogenesis assay. Jestin George is a research technician working on the LRG1 project in the Department of Cell Biology, University College London Institute of Ophthalmology, London, UK.
For more information on Xiaomeng WANG’s lab, please click here.
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