The Chorioallantoic Membrane (CAM) Assay for the Analysis of Starvation-Induced Autophagy.

Q4 Biochemistry, Genetics and Molecular Biology
Nazlı Şevval Menemenli, Özün Özcan, H Hazal Hüsnügil, Aliye Ezgi Güleç Taşkıran, Göksu Oral, Aytekin Akyol, Sreeparna Banerjee
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引用次数: 0

Abstract

During avian development, the chorioallantoic membrane (CAM) is generated around 4 days after fertilization following the fusion of the allantois and the chorion. The CAM develops rapidly over the next several days and gets heavily vascularized and therefore has been explored widely as a tool for the study of angiogenesis. Additionally, being immunodeficient, the CAM can be used for tumor growth of human origin and its metastasis. Of note, the CAM assay is minimally invasive for the chicken embryo and lacks innervation, which gives this in vivo model a low ethical burden. Here, we describe the protocol for the generation of microtumors from human colorectal cancer cell lines on the CAM, incubated in a nutrient-deficient medium for the activation of autophagy. We show that pre-inoculation markers of autophagy induced through nutrient deficiency are retained in the microtumors generated on the CAM.

用于分析饥饿诱导的自噬的绒毛膜 (CAM) 分析法。
在鸟类的发育过程中,绒毛膜(CAM)是在受精后 4 天左右由尿囊和绒毛膜融合产生的。绒毛膜在接下来的几天内迅速发育,并形成大量血管,因此被广泛用作研究血管生成的工具。此外,CAM 免疫缺陷,可用于人类肿瘤的生长和转移。值得注意的是,CAM 试验对鸡胚胎的创伤很小,而且缺乏神经支配,因此这种体内模型的伦理负担很低。在此,我们介绍了在 CAM 上用人类结直肠癌细胞系生成微瘤的方案,该微瘤在营养缺乏的培养基中培养,以激活自噬。我们的研究表明,在 CAM 上生成的微瘤中保留了通过营养缺乏诱导自噬的接种前标记。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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