The fetal mouse metatarsal bone explant as a model of angiogenesis.
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.
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
Weihua
W
Song
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
Chee Wai
CW
Fhu
Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Proteos, Singapore, Singapore.
Koon Hwee
KH
Ang
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
Cheng Hao
CH
Liu
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
Nurul Azizah Binte
NA
Johari
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
Daniel
D
Lio
Institute of Cardiovascular Sciences, Manchester, UK.
Sabu
http://orcid.org/0000-0002-0916-5827
S
Abraham
Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Proteos, Singapore, Singapore.
Wanjin
W
Hong
Department of Cell Biology, Institute of Ophthalmology, University College London, London, UK.
Stephen E
SE
Moss
Department of Cell Biology, Institute of Ophthalmology, University College London, London, UK.
John
J
Greenwood
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.|Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Proteos, Singapore, Singapore.|Department of Cell Biology, Institute of Ophthalmology, University College London, London, UK.
Xiaomeng
X
Wang
D000818:Animals; D046508:Culture Techniques / Q000379:methods*; D004195:Disease Models, Animal; D005260:Female; D005333:Fetus; D005455:Fluorescent Antibody Technique; D008682:Metatarsal Bones / Q000098:blood supply* / Q000473:pathology; D051379:Mice; D055806:Microvessels / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000473:pathology*; D013194:Staining and Labeling