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.
摘要：
在鸟类发育过程中,尿囊和绒毛膜融合后受精后约4天产生绒毛尿囊膜(CAM)。CAM在接下来的几天内迅速发展，并大量血管化，因此已被广泛用作研究血管生成的工具。此外,免疫缺陷，CAM可用于人类起源的肿瘤生长及其转移。值得注意的是，CAM测定对鸡胚的侵入性最小，缺乏神经支配，这给了这个体内模型一个低的道德负担。这里,我们描述了在CAM上从人类结直肠癌细胞系产生微肿瘤的协议，在营养缺乏的培养基中孵育以激活自噬。我们表明，通过营养缺乏诱导的自噬的接种前标志物保留在CAM上产生的微肿瘤中。
