Chick chorioallantoic membrane model to investigate role of migrasome in angiogenensis.

Biophysics reports Pub Date : 2023-10-31 DOI:10.52601/bpr.2023.230021

Cuifang Zhang, Helen He, Shuyao Yin, Mingyi Gao, Li Yu

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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.

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用小鸡绒毛膜模型研究移行体在血管生成中的作用

血管系统的发育对胚胎发育至关重要，其中包括血管生成等过程。血管生成在许多正常的生理和病理过程中发挥着关键作用。它由一系列血管生成蛋白驱动，包括血管生成生长因子、趋化因子和细胞外基质蛋白。在各种动物模型系统中，绒毛膜（CAM）已被证明是分析候选细胞或因子血管生成潜能的重要工具。在本方案中，我们将提供建立 CAM 模型的详细步骤，以评估迁移体在胚胎血管生成中的功能和机制。其中包括评估CAM血管生成的CAM尼龙网试验和CAM体内外发芽试验，以及移行体的观察、纯化和输送。此外，我们还介绍了利用CRISPR系统在CAM组织内生成T4-KO-mCherry-KI胚胎，以研究移行体在血管生成中的作用。

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Biophysics reports

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8 weeks