Three-Dimensional Culture Decreases the Angiogenic Ability of Mouse Macrophages

Macrophages play important roles in angiogenesis; however, previous studies on macrophage angiogenesis have focused on traditional 2D cultures. In this study, we established a 3D culture system for macrophages using collagen microcarriers and assessed the effect of 3D culture on their angiogenic capabilities. Macrophages grown in 3D culture displayed a significantly different morphology and arrangement under electron microscopy compared to those grown in 2D culture. Tube formation assays and chick embryo chorioallantoic membrane assays further revealed that 3D-cultured macrophages were less angiogenic than those in 2D culture. Whole-transcriptome sequencing showed that nearly 40% of genes were significantly differently expressed, including nine important angiogenic factors of which seven had been downregulated. In addition, the expression of almost all genes related to two important angiogenic pathways was decreased in 3D-cultured macrophages, including the two key angiogenic factors, VEGFA and ANG2. Together, the findings of our study improve our understanding of angiogenesis and 3D macrophage culture in tissues, and provide new avenues and methods for future research on macrophages.

Applications of the Chick Chorioallantoic Membrane as an Alternative Model for Cancer Studies

A variety of in vivo experimental models have been established for the studies of human cancer using both cancer cell lines and patient-derived xenografts (PDXs). In order to meet the aspiration of precision medicine, the in vivomurine models have been widely adopted. However, common constraints such as high cost, long duration of experiments, and low engraftment efficiency remained to be resolved. The chick embryo chorioallantoic membrane (CAM) is an alternative model to overcome some of these limitations. Here, we provide an overview of the applications of the chick CAM model in the study of oncology. The CAM model has shown significant retention of tumor heterogeneity alongside increased xenograft take rates in several PDX studies. Various imaging techniques and data analysis have been applied to study tumor metastasis, angiogenesis, and therapeutic response to novel agents. Lastly, to practically illustrate the feasibility of utilizing the CAM model, we summarize the general protocol used in a case study utilizing an ovarian cancer PDX.

Utilisation of Chick Embryo Chorioallantoic Membrane as a Model Platform for Imaging-Navigated Biomedical Research

The fertilised chick egg and particularly its chorioallantoic membrane (CAM) have drawn continuing interest in biomedicine and bioengineering fields, especially for research on vascular study, cancer, drug screening and development, cell factors, stem cells, etc. This literature review systemically introduces the CAM’s structural evolution, functions, vascular features and the circulation system, and cell regulatory factors. It also presents the major and updated applications of the CAM in assays for pharmacokinetics and biodistribution, drug efficacy and toxicology testing/screening in preclinical pharmacological research. The time course of CAM applications for different assays and their advantages and limitations are summarised. Among these applications, two aspects are emphasised: (1) potential utility of the CAM for preclinical studies on vascular-disrupting agents (VDAs), promising for anti-cancer vascular-targeted therapy, and (2) modern imaging technologies, including modalities and their applications for real-time visualisation, monitoring and evaluation of the changes in CAM vasculature as well as the interactions occurring after introducing the tested medical, pharmaceutical and biological agents into the system. The aim of this article is to help those working in the biomedical field to familiarise themselves with the chick embryo CAM as an alternative platform and to utilise it to design and optimise experimental settings for their specific research topics.

Preparation, Preliminary Characterization and Antiangiogenic Activities of Polysaccharides from Pomegranate Peels

In the present study, the crude polysaccharides from pomegranate peels (CPP) were prepared by compound enzyme-extraction technology, and the response surface methodology was used to optimize the extraction parameters. Box–Behnken design (BBD) was applied to estimate the effects of extraction temperature, extraction pH, and dosage of compound enzyme on the yield of CPP. A mathematical model with high fitness was obtained. Extraction temperature, extraction pH, and dosage of compound enzyme exhibited independent and interactive effects on CPP yield. CPP were purified by macroporous resin HP-20 and Sephadex G-100 column chromatography to afford purified fraction of CPP-2. The relative molecular weight of CPP-2 was 93.5 kDa. In CPP-2, and FT-IR spectra showed that the main components among CPP-2 may be pectic polysaccharides.The effect of CPP-2 on angiogenesis was measured in vivo by using the chick embryo chorioallantoic membrane (CAM) assay. The results demonstrated that CPP-2 suppressed angiogenesis in chicken embryos.

Spatial distribution of blood vessels in the chick embryo chorioallantoic membrane

The chick embryo chorioallantoic membrane (CAM) is a useful tool to study both angiogenesis and anti-angiogenesis in vivo. CAM vascular growth pattern including the way through vessels fills the available space can be pretty easily described and quantified using image analysis procedures, in order to evaluate different parameters, including fractal dimension, lacunarity and non-fractal order-disorder parameters. In the present study, we further expanded this morphological description, by estimating an index expressing the degree of symmetry characterizing the CAM vascular tree structure in the course of the embryonic development. Moreover, a uniformity index was estimated to quantitatively characterize the space-filling features of the vessels, i.e. the degree of spatial uniformity of their distribution in the tissue.

Exploitation of the chick embryo chorioallantoic membrane (CAM) as a platform for anti-metastatic drug testing

The establishment of clinically relevant models for tumor metastasis and drug testing is a major challenge in cancer research. Here we report a physiologically relevant assay enabling quantitative analysis of metastatic capacity of tumor cells following implantation into the chorioallantoic membrane (CAM). Engraftment of as few as 103 non-small cell lung cancer (NSCLC) and prostate cancer (PCa) cell lines was sufficient for both primary tumor and metastasis formation. Standard 2D-imaging as well as 3D optical tomography imaging were used for the detection of fluorescent metastatic foci in the chick embryo. H2228- and H1975-initiated metastases were confirmed by genomic analysis. We quantified the inhibitory effect of docetaxel on LNCaP, and that of cisplatin on A549- and H1299-initiated metastatic growths. The CAM assay also mimicked the sensitivity of ALK-rearranged H2228 and EGFR-mutated H1975 NSCLC cells to tyrosine kinase inhibitors crizotinib and gefitinib respectively, as well as sensitivity of LNCaP cells to androgen-dependent enzalutamide therapy. The assay was suggested to reconstitute the bone metastatic tropism of PCa cells. We show that the CAM chick embryo model may be a powerful preclinical platform for testing and targeting of the metastatic capacity of cancer cells.