Chick chorioallantoic membrane model to investigate role of migrasome in angiogenensis

Abstract

The development of the vascular system is essential for embryonic development, including processes such as angiogenesis. Angiogenesis plays a critical role in many normal physiological and pathological processes. It is driven by a set of angiogenic proteins, including angiogenic growth factors, chemokines, and extracellular matrix proteins. Among various animal model systems, the chorioallantoic membrane (CAM), a specialized and highly vascularized tissue of the avian embryo, has proven to be a valuable tool for analyzing the angiogenic potential of candidate cells or factors. In this protocol, we provide detailed procedures for establishing the CAM model to evaluate the function and mechanism of migrasomes in embryonic angiogenesis. This includes the CAM nylon mesh assay and CAM ex vivo sprouting assay to assess CAM angiogenesis, as well as the observation, purification, and delivery of migrasomes. Additionally, we describe the generation of T4-KO-mCherry-KI embryos using the CRISPR system within the CAM tissue to investigate the role of migrasomes in angiogenesis.

Graphical abstract

Keywords

Angiogenesis / Chorioallantoic membrane (CAM) / Chick CAM model

Acknowledgements

We are grateful to the members of the Yu groups for their helpful discussions and suggestions. We are grateful to the members of the Zhihua Liu groups for their helpful discussions and suggestions about the CRISPR-Cas9 system. This research was supported by the Ministry of Science and Technology of the People’s Republic of China (grant no. 2017YFA0503404), the National Natural Science Foundation of China (31790401, 92054301 and 32030023), and the Beijing Municipal Science & Technology Commission (Z201100005320019 and Z211100003321002) and internal grant from Tsinghua university (2030 plan). We thank the State Key Laboratory of Membrane Biology for facility support and assistance with spinning disk microscopy. We would like to acknowledge the assistance of SLSTU-Nikon Biological Imaging Center for assistance with using the NIKON A1RSiHD25 laser scanning confocal microscope and data analysis by NLS element software.

RIGHTS & PERMISSIONS

2023 The Author(s)