scholarly journals Optimization of the chicken chorioallantoic membrane assay as reliable in vivo model for the analysis of osteosarcoma

PLoS ONE ◽  
2019 ◽  
Vol 14 (4) ◽  
pp. e0215312 ◽  
Author(s):  
Pierre Kunz ◽  
Astrid Schenker ◽  
Heiner Sähr ◽  
Burkhard Lehner ◽  
Jörg Fellenberg
Keyword(s):  
Chorioallantoic Membrane ◽  
In Vivo Model ◽  
Chicken Chorioallantoic Membrane ◽  
Chorioallantoic Membrane Assay

scholarly journals Novel tempeh (fermented soyabean) isoflavones inhibit in vivo angiogenesis in the chicken chorioallantoic membrane assay

2005 ◽  
Vol 93 (3) ◽  
pp. 317-323 ◽  
Author(s):  
Serafim Kiriakidis ◽  
Oliver Högemeier ◽  
Susanne Starcke ◽  
Frank Dombrowski ◽  
Jens Claus Hahne ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cell ◽  
Molecular Mechanisms ◽  
Inflammatory Diseases ◽  
Chorioallantoic Membrane ◽  
Endothelial Cell Proliferation ◽  
Inhibition Of Angiogenesis ◽  
Chicken Chorioallantoic Membrane ◽  
Chorioallantoic Membrane Assay

Anti-angiogenic strategies are emerging as an important tool for the treatment of cancer and inflammatory diseases. In the present investigation we isolated several isoflavones from a tempeh (fermented soyabean) extract. The isolated isoflavones were identified as 5,7,4′-trihydroxyisoflavone (genistein), 7,4′-dihydroxyisoflavone (daidzein), 6,7,4′-trihydroxyisoflavone (factor 2), 7,8,4′-trihydroxyisoflavone (7,8,4′-TriOH) and 5,7,3′,4′-tetrahydroxyisoflavone (orobol). The effects on angiogenesis of these isoflavones were evaluated in the chicken chorioallantoic membrane assay; their capacity to inhibit vascular endothelial growth factor-induced endothelial cell proliferation and expression of the Ets 1 transcription factor, known to be implicated in the regulation of new blood vessel formation, were also investigated. We found that all isoflavones inhibited angiogenesis, albeit with different potencies. Compared with negative controls, which slightly inhibited in vivo angiogenesis by 6·30 %, genistein reduced angiogensis by 75·09 %, followed by orobol (67·96 %), factor 2 (56·77 %), daidzein (48·98 %) and 7,8,4′-TriOH (24·42 %). These compounds also inhibited endothelial cell proliferation, with orobol causing the greatest inhibition at lower concentrations. The isoflavones also inhibited Ets 1 expression, providing some insight into the molecular mechanisms of their action. Furthermore, the chemical structure of the different isoflavones suggests a structure–activity relationship. Our present findings suggest that the new isoflavones might be added to the list of low molecular mass therapeutic agents for the inhibition of angiogenesis.

scholarly journals QUANTIFICATION OF ANGIOGENESIS IN THE CHICKEN CHORIOALLANTOIC MEMBRANE (CAM)

2011 ◽  
Vol 23 (3) ◽  
pp. 169 ◽  
Author(s):  
Silvia Blacher ◽  
Laetitia Devy ◽  
Ruslan Hlushchuk ◽  
Etienne Larger ◽  
Noel Lamandé ◽  
...  
Keyword(s):  
Chorioallantoic Membrane ◽  
In Vivo Model ◽  
New Developments ◽  
Chick Chorioallantoic Membrane ◽  
Capillary Plexus ◽  
Feeding Vessels ◽  
Two Phases ◽  
Chicken Chorioallantoic Membrane ◽  
Accurate Quantification

The chick chorioallantoic membrane (CAM) provides a suitable in vivo model to study angiogenesis and evaluate several pro- and anti-angiogenic factors and compounds. In the present work, new developments in image analysis are used to quantify CAM angiogenic response from optical microscopic observations, covering all vascular components, from the large supplying and feeding vessels down to the capillary plexus. To validate our methodology angiogenesis is quantified during two phases of CAM development (day 7 and 13) and after treatment with an antiangiogenic modulator of the angiogenesis. Our morphometric analysis emphasizes that an accurate quantification of the CAM vasculature needs to be performed at various scales.

scholarly journals In Situ DNA/Protein Interaction Assay to Visualize Transcriptional Factor Activation

2020 ◽  
Vol 3 (4) ◽  
pp. 80
Author(s):  
Michela Corsini ◽  
Emanuela Moroni ◽  
Cosetta Ravelli ◽  
Elisabetta Grillo ◽  
Marco Presta ◽  
...  
Keyword(s):  
Protein Interaction ◽  
Nuclear Translocation ◽  
Chorioallantoic Membrane ◽  
Camp Response Element Binding ◽  
Transcriptional Factors ◽  
In Vivo Model ◽  
Element Binding Protein ◽  
Dna Protein Interaction

The chick embryo chorioallantoic membrane (CAM) represents a powerful in vivo model to study several physiological and pathological processes including inflammation and tumor progression. Nevertheless, the possibility of deepening the molecular processes in the CAM system is biased by the absence/scarcity of chemical and biological reagents, designed explicitly for avian species. This is particularly true for transcriptional factors, proteinaceous molecules that regulate various cellular responses, including proliferation, survival, and differentiation. Here, we propose a detailed antibody-independent protocol to visualize the activation and nuclear translocation of transcriptional factors in cells or in tissues of different animal species. As a proof of concept, DNA/cAMP response element-binding protein (CREB) interaction was characterized on the CAM tissue using oligonucleotides containing the palindromic binding sequence of CREB. Scrambled oligonucleotides were used as controls. In situ DNA/protein interaction protocol is a versatile method that is useful for the study of transcription factors in the cell and tissue of different origins.

scholarly journals Degradation of Mitochondria and Oxidative Stress as the Main Mechanism of Toxicity of Pristine Graphene on U87 Glioblastoma Cells and Tumors and HS-5 Cells

2019 ◽  
Vol 20 (3) ◽  
pp. 650 ◽  
Author(s):  
Sławomir Jaworski ◽  
Barbara Strojny ◽  
Ewa Sawosz ◽  
Mateusz Wierzbicki ◽  
Marta Grodzik ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Chorioallantoic Membrane ◽  
In Vivo Model ◽  
Pristine Graphene ◽  
Infrared Measurements ◽  
The Impact

Due to the development of nanotechnologies, graphene and graphene-based nanomaterials have attracted immense scientific interest owing to their extraordinary properties. Graphene can be used in many fields, including biomedicine. To date, little is known about the impact graphene may have on human health in the case of intentional exposure. The present study was carried out on U87 glioma cells and non-cancer HS-5 cell lines as in vitro model and U87 tumors cultured on chicken embryo chorioallantoic membrane as in vivo model, on which the effects of pristine graphene platelets (GPs) were evaluated. The investigation consisted of structural analysis of GPs using transmission electron microscopy, Fourier transmission infrared measurements, zeta potential measurements, evaluation of cell morphology, assessment of cell viability, investigation of reactive oxygen species production, and investigation of mitochondrial membrane potential. The toxicity of U87 glioma tumors was evaluated by calculating the weight and volume of tumors and performing analyses of the ultrastructure, histology, and protein expression. The in vitro results indicate that GPs have dose-dependent cytotoxicity via ROS overproduction and depletion of the mitochondrial membrane potential. The mass and volume of tumors were reduced in vivo after injection of GPs. Additionally, the level of apoptotic and necrotic markers increased in GPs-treated tumors.

scholarly journals In vivo paracrine interaction between urokinase and its receptor: effect on tumor cell invasion.

1991 ◽  
Vol 115 (4) ◽  
pp. 1107-1112 ◽  
Author(s):  
L Ossowski ◽  
G Clunie ◽  
M T Masucci ◽  
F Blasi
Keyword(s):  
Chorioallantoic Membrane ◽  
Fold Increase ◽  
Cell Types ◽  
Cell Surface Receptor ◽  
In Vivo Model ◽  
Specific Cell ◽  
Upa Receptor ◽  
Surface Receptor

Numerous studies have linked the production of increased levels of urokinase type plasminogen activator (uPA) with the malignant phenotype. It has also been shown that a specific cell surface receptor can bind uPA through a domain distinct and distant from the proteolytic domain. In an in vivo model of invasion, consisting of experimentally modified chorioallantoic membrane (CAM) of a chick embryo, only cells that concurrently expressed both uPA and a receptor for uPA, and in which the receptor was saturated with uPA, were efficient in invasion. To test whether uPA produced by one cell can, in a paracrine fashion, affect the invasive capacity of a receptor-expressing cell, we transfected LB6 mouse cells with human uPA (LB6[uPA]), or human uPA-receptor cDNA (LB6[uPAR]). LB6(uPA) cells released into the medium 1-2 Ploug units of human uPA per 10(6) cells in 24 h. The LB6(uPAR) cells expressed on their surface approximately 12,000 high affinity (Kd 1.7 x 10(-10) M uPA binding sites per cell. Unlabeled LB6(uPA) and 125-IUdR-labeled LB6(uPAR) cells were coinoculated onto experimentally wounded and resealed CAMs and their invasion was compared to that of homologous mixtures of labeled and unlabeled LB6(uPAR) or LB6(uPA) cells. Concurrent presence of both cell types in the CAMs resulted in a 1.8-fold increase of invasion of the uPA-receptor expressing cells. A four-fold stimulation of invasion was observed when cells were cocultured in vitro, prior to in vivo inoculation. Enhancement of invasion was prevented in both sets of experiments by treatment with specific antihuman uPA antibodies, indicating that uPA was the main mediator of the invasion-enhancing, paracrine effect on the receptor-expressing cells.

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