Abstract
The chick chorioallantoic membrane (CAM) assay model of angiogenesis has been highlighted as a relatively quick, low cost and effective model for the study of pro-angiogenic and anti-angiogenic factors. The chick CAM is a highly vascularised extraembryonic membrane which functions for gas exchange, nutrient exchange and waste removal for the growing chick embryo. It is beneficial as it can function as a treatment screening tool, which bridges the gap between cell based in vitro studies and in vivo animal experimentation. In this review, we explore the benefits and drawbacks of the CAM assay to study microcirculation, by the investigation of each distinct stage of the CAM assay procedure, including cultivation techniques, treatment applications and methods of determining an angiogenic response using this assay. We detail the angiogenic effect of treatments, including drugs, metabolites, genes and cells used in conjunction with the CAM assay, while also highlighting the testing of genetically modified cells. We also present a detailed exploration of the advantages and limitations of different CAM analysis techniques, including visual assessment, histological and molecular analysis along with vascular casting methods and live blood flow observations.
Highlights
The appropriate delivery of metabolites and removal of waste products is essential in maintaining tissue homeostasis in the body
Angiogenesis is the process by which new blood vessels form from pre-existing vessels, a phenomenon required in normal physiology, development, growth injury and disease [3]
Zhang et al used an expression plasmid to inhibit the anti-angiogenic function of miR-338 in hepatocellular carcinoma, leading to a significant increase in small blood vessel formation in the chorioallantoic membrane (CAM) assay [194]
Summary
The appropriate delivery of metabolites and removal of waste products is essential in maintaining tissue homeostasis in the body. For this to occur, the presence of a vast well-connected microvascular blood vessel network is crucial. Angiogenesis is the process by which new blood vessels form from pre-existing vessels, a phenomenon required in normal physiology, development, growth injury and disease [3]. Intussusceptive angiogenesis was more recently discovered and entails pre-existing blood vessels “splitting” down the middle to form two new branching blood vessels [6]. Angiogenesis is an organised cascade of events, regulated by several pro- and anti-angiogenic growth factors. The growth of new blood vessels is induced by the delicate balance between pro-angiogenic and anti-angiogenic factors [16].
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