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Domenico Ribatti - One of the best experts on this subject based on the ideXlab platform.
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the chick embryo chorioallantoic membrane
In Vivo Models to Study Angiogenesis, 2018Co-Authors: Domenico RibattiAbstract:The chick embryo chorioallantoic membrane (CAM) is an extraembryonic membrane which serves as a gas exchange surface and its function is supported by a dense capillary network. Because of its extensive vascularization and easy accessibility, the CAM has been broadly used to study the morphofunctional aspects of the angiogenesis process in vivo and to investigate the efficacy and mechanisms of action of proangiogenic and antiangiogenic natural and synthetic molecules. The CAM is a favored system for the study of tumor angiogenesis and metastasis, because at this stage the chick immunocompetence system is not fully developed and the conditions for rejection have not been established. Tumors remain avascular for 72 h, after which they are penetrated by new blood vessels and begin a phase of rapid growth. The rate of growth during this vascular phase is greater for implants on days 5–6 and decreases for later days of implantation. Other studies using the tumor cells/CAM model have focused on the invasion of the chorionic epithelium and the blood vessels by tumor cells. The cells invade the epithelium and the mesenchymal connective tissue below, where they are found in the form of a dense bed of blood vessels, which is a target for intravasation.
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the chick embryo chorioallantoic membrane cam assay
Reproductive Toxicology, 2017Co-Authors: Domenico RibattiAbstract:The chick embryo chorioallantoic membrane (CAM) is an extraembryonic membrane which serves as a gas exchange surface and its function is supported by a dense capillary network. Because of its extensive vascularization and easy accessibility, the CAM has been used to study the morphofunctional aspects of the angiogenesis process in vivo and to investigate the efficacy and mechanisms of action of pro-angiogenic and anti-angiogenic natural molecules. The CAM has long been a favored system for the study of tumor angiogenesis and metastasis, because a this stage the chick immunocompetence system is not fully developed and the conditions for rejection have not been established.
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the chick embryo chorioallantoic membrane as a model for tumor biology
Experimental Cell Research, 2014Co-Authors: Domenico RibattiAbstract:Abstract Among the in vivo models, the chick embryo chorioallantoic membrane (CAM) has been used to implant several tumor types as well as malignant cell lines to study their growth rate, angiogenic potential and metastatic capability. This review article is focused on the major compelling literature data on the use of the CAM to investigate tumor growth and the metastatic process.
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the chick embryo chorioallantoic membrane in the study of angiogenesis and metastasis the cam assay in the study of angiogenesis and metastasis
2010Co-Authors: Domenico RibattiAbstract:1. Introduction 2. Chorioallantoic membrane and its embryological origin 2.1. Morphology of chorioallantoic membrane blood and lymphatic vessels 2.2. A single blood sinus or a capillary plexus beneath the chorionic epithelium? 2.3. The chorioallantoic membrane vascular growth 3. Use of chorioallantoic membrane in the study of angiogenic molecules 3.1. Role of FGF-2 in chorioallantoic membrane vascularization 4. Use of chorioallantoic membrane in the study of antiangiogenic molecules 5. Use of chorioallantoic membranes in the study of tumor angiogenesis 5.1. Angiogenesis and antiangiogenesis in multiple myeloma 5.2. Angiogenesis and antiangiogenesis in human neuroblastoma 6. Use of chorioallantoic membrane in the study of tumor metastasis 6.1. Spontaneous metastasis models. 6.2. Experimental metastasis studies. 7. Other applications 7.1. In ovo and ex ovo methods 7.2. Use of chorioallantoic membrane in the study of tumor lymphangiogenesis 8. Different morphological techniques that can be used to study vascularization of the CAM and the genes involved. 9. Methods of quantifying the angiogenic response 10. Limitations of the chorioallantoic membrane assay 11. Other classical in vivo assays in comparison (advantages, disadvantages and limitations) with the chorioallantoic membrane 12. Concluding remarks Acknowledgments References
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the chick embryo chorioallantoic membrane in the study of tumor angiogenesis
Romanian Journal of Morphology and Embryology, 2008Co-Authors: Domenico RibattiAbstract:: The chick embryo chorioallantoic membrane (CAM) is commonly used as an experimental in vivo assay to study both angiogenesis and antiangiogenesis in response to tissues, cells or soluble factors. This article summarizes literature data about the use of the CAM in the study of tumor angiogenesis and particularly our experimental data concerning the study of angiogenesis in multiple myeloma and in neuroblastoma.
Marco Presta - One of the best experts on this subject based on the ideXlab platform.
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the gelatin sponge chorioallantoic membrane assay
Nature Protocols, 2006Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Marco PrestaAbstract:Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge–CAM assay is simple, inexpensive and suitable for large-scale screening. *Note: In the version of the article initially published online, references 6 and 7 were incorrect. The correct references are: 6. Serbedzija, G.N., Flynn, E. & Willet, C.E. Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3, 519–528 (2000). 7. Ribatti, D., Vacca, A., Roncali, L. & Dammacco, F. The chick embryo chorioallantoic membrane as a model for in vivo research on angiogenesis. Int. J. Dev. Biol. 40, 1189–1897 (1996). The error has been corrected in all versions of the article.
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new model for the study of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane the gelatin sponge chorioallantoic membrane assay
Journal of Vascular Research, 1997Co-Authors: Domenico Ribatti, Angelo Vacca, Luisa Roncali, Anna Gualandris, Maria Bastaki, Monica Iurlaro, Marco PrestaAbstract:Several methods for the in vivo study of angiogenesis are available, and each angiogenic assay presents distinct advantages and disadvantages. In this study, we present a new method for the quantitati
Angelo Vacca - One of the best experts on this subject based on the ideXlab platform.
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the gelatin sponge chorioallantoic membrane assay
Nature Protocols, 2006Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Marco PrestaAbstract:Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge–CAM assay is simple, inexpensive and suitable for large-scale screening. *Note: In the version of the article initially published online, references 6 and 7 were incorrect. The correct references are: 6. Serbedzija, G.N., Flynn, E. & Willet, C.E. Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3, 519–528 (2000). 7. Ribatti, D., Vacca, A., Roncali, L. & Dammacco, F. The chick embryo chorioallantoic membrane as a model for in vivo research on angiogenesis. Int. J. Dev. Biol. 40, 1189–1897 (1996). The error has been corrected in all versions of the article.
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inhibition of endothelial cell functions and of angiogenesis by the metastasis inhibitor nami a
British Journal of Cancer, 2002Co-Authors: Angelo Vacca, Domenico Ribatti, M Bruno, Angelina Boccarelli, Mauro Coluccia, Alberta Bergamo, Spiridione Garbisa, Luigi Sartor, Gianni SavaAbstract:NAMI-A is a ruthenium-based compound with selective anti-metastasis activity in experimental models of solid tumours. We studied whether this activity was dependent on anti-angiogenic ability of NAMI-A. We thus investigated its in vitro effects on endothelial cell functions necessary for angiogenesis to develop, as well as its in vivo effects in the chick embryo chorioallantoic membrane model. Endothelial cell proliferation, chemotaxis, and secretion of the matrix-degrading enzyme metalloproteinase-2 were inhibited by NAMI-A in a dose-dependent manner, and without morphologic signs of cell apoptosis or necrosis. Lastly, NAMI-A displayed a dose-dependent in vivo anti-angiogenic activity in the chorioallantoic membrane model. These data suggest that the anti-angiogenic activity of NAMI-A can contribute to its anti-metastatic efficacy in mice bearing malignant solid tumours.
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mast cells and their secretory granules are angiogenic in the chick embryo chorioallantoic membrane
Clinical & Experimental Allergy, 2001Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Luisa Roncali, Enrico Crivellato, Luigi Candussio, F DammaccoAbstract:Background Many data suggest that the density of mast cells is highly correlated with the extent of both normal and pathological angiogenesis. Objective In this study we have compared in an in vivo assay, the chick embryo chorioallantoic membrane, the angiogenic potential of mast cell suspensions isolated from rats, degranulated mast cells and their secretory granules. Methods Gelatin sponges adsorbed with cell suspensions of rat mast cells, degranulated mast cells and their secretory granules were implanted on the top of the chorioallantoic membrane at day 8 of incubation. At day 12 the angiogenic response was evaluated macroscopically, microscopically and by a morphometric method of ‘point counting’. Results Isolated mast cells and their secretory granules, but not degranulated mast cells, induced an angiogenic response in the chorioallantoic membrane. The addition of antifibroblast growth factor-2 or antivascular endothelial growth factor antibodies reduced the angiogenic response of both mast cells and their secretory granules by 50% and 30%, respectively. Conclusion These data support the evidence that the angiogenic properties of mast cells depend on the angiogenic molecules contained in their secretory granules and indicate that fibroblast growth factor-2 and vascular endothelial growth factor are the angiogenic cytokines primarily and perhaps synergistically responsible for this vasoproliferative activity.
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new model for the study of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane the gelatin sponge chorioallantoic membrane assay
Journal of Vascular Research, 1997Co-Authors: Domenico Ribatti, Angelo Vacca, Luisa Roncali, Anna Gualandris, Maria Bastaki, Monica Iurlaro, Marco PrestaAbstract:Several methods for the in vivo study of angiogenesis are available, and each angiogenic assay presents distinct advantages and disadvantages. In this study, we present a new method for the quantitati
Beatrice Nico - One of the best experts on this subject based on the ideXlab platform.
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the gelatin sponge chorioallantoic membrane assay
Nature Protocols, 2006Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Marco PrestaAbstract:Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge–CAM assay is simple, inexpensive and suitable for large-scale screening. *Note: In the version of the article initially published online, references 6 and 7 were incorrect. The correct references are: 6. Serbedzija, G.N., Flynn, E. & Willet, C.E. Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3, 519–528 (2000). 7. Ribatti, D., Vacca, A., Roncali, L. & Dammacco, F. The chick embryo chorioallantoic membrane as a model for in vivo research on angiogenesis. Int. J. Dev. Biol. 40, 1189–1897 (1996). The error has been corrected in all versions of the article.
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aquaporin 1 expression in the chick embryo chorioallantoic membrane
Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology, 2002Co-Authors: Domenico Ribatti, Beatrice Nico, Luisa Roncali, Antonio Frigeri, Grazia Paola Nicchia, Michela De Giorgis, Maria SveltoAbstract:The chick embryo chorioallantoic membrane (CAM) is commonly used in vivo to study both angiogenesis and anti-angiogenesis. Rapid membrane water transport is mediated by a family of molecular water channels, called aquaporins (AQPs), which have been identified in the epithelial and endothelial cells of higher vertebrates. AQP1, expressed in adsorptive and secretory epithelia, is also expressed in endothelial cells of capillaries and arteries. Its mRNA has been found in vascular smooth muscle cells (VSMCs) of arteries and capillaries, as well as in a subset of VSMCs of human atherosclerotic plaques. This study investigated the developmental expression of AQP1 in the chick CAM by Western blot and immunohistochemistry. Western blot results show that a major nonglycosylated band was observed with electrophoretic mobility of approximately 28 kDa in the three developmental stages examined. Immunohistochemistry data demonstrate that AQP1 was clearly expressed in the ectodermal and endodermal epithelia, the vascular endothelium, and the VSMCs. Because little information is available on the behavior of microvessel AQP1 during angiogenesis in normal and pathological conditions, our data relative to the pattern of expression of AQP1 in CAM blood vessels in normal conditions may be considered a useful tool to further investigate its modifications in several experimental conditions implying a stimulation or an inhibition of angiogenesis in the CAM assay. Anat Rec 268:85–89, 2002. © 2002 Wiley-Liss, Inc.
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mast cells and their secretory granules are angiogenic in the chick embryo chorioallantoic membrane
Clinical & Experimental Allergy, 2001Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Luisa Roncali, Enrico Crivellato, Luigi Candussio, F DammaccoAbstract:Background Many data suggest that the density of mast cells is highly correlated with the extent of both normal and pathological angiogenesis. Objective In this study we have compared in an in vivo assay, the chick embryo chorioallantoic membrane, the angiogenic potential of mast cell suspensions isolated from rats, degranulated mast cells and their secretory granules. Methods Gelatin sponges adsorbed with cell suspensions of rat mast cells, degranulated mast cells and their secretory granules were implanted on the top of the chorioallantoic membrane at day 8 of incubation. At day 12 the angiogenic response was evaluated macroscopically, microscopically and by a morphometric method of ‘point counting’. Results Isolated mast cells and their secretory granules, but not degranulated mast cells, induced an angiogenic response in the chorioallantoic membrane. The addition of antifibroblast growth factor-2 or antivascular endothelial growth factor antibodies reduced the angiogenic response of both mast cells and their secretory granules by 50% and 30%, respectively. Conclusion These data support the evidence that the angiogenic properties of mast cells depend on the angiogenic molecules contained in their secretory granules and indicate that fibroblast growth factor-2 and vascular endothelial growth factor are the angiogenic cytokines primarily and perhaps synergistically responsible for this vasoproliferative activity.
Michael B Thompson - One of the best experts on this subject based on the ideXlab platform.
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changes to the uterine epithelium during the reproductive cycle of two viviparous lizard species niveoscincus spp
Acta Zoologica, 2015Co-Authors: Qiong Wu, Michael B Thompson, Cameron K Fong, Christopher R MurphyAbstract:We investigated morphological differences in uterine epithelia of the reproductive cycle between two closely related viviparous skinks, Niveoscincus metallicus (lecithotrophic) and Niveoscincus ocellatus (placentotrophic), which have similar placental complexity but different degrees of placentotrophy. Scanning (SEM) and transmission electron microscopy (TEM) revealed that the uterine surface of non-reproductive females of both species is mainly covered by ciliated cells. As vitellogenesis begins, the uterine epithelium consists of ciliated and non-ciliated cells under a thin glycocalyx. Microvilli are greatly reduced at mid-pregnancy, and the uterus differentiates into two structurally distinct regions: the chorioallantoic and the omphaloplacenta. At late stages of pregnancy, the uterine epithelium of chorioallantoic placenta in both species is further ridged, forming a knobbly uterine surface. The ultrastructural evidence between N. metallicus and N. ocellatus cannot strictly account for the distinct differences in their placentotrophy; as yet unexplored molecular nutrient transport mechanisms that are not reflected in uterine ultrastructure must play significant roles in nutrient transportation. Characteristics consistent with a plasma membrane transformation were confirmed in both species.
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uterine and chorioallantoic angiogenesis and changes in the uterine epithelium during gestation in the viviparous lizard niveoscincus conventryi squamata scincidae
Journal of Morphology, 2012Co-Authors: Martha Patricia Ramirezpinilla, Scott L Parker, Christopher R Murphy, Michael B ThompsonAbstract:We used immunofluorescent confocal microscopy and scanning electron microscopy to quantify uterine vascularity and to describe uterine surface morphology during gestation in pregnant females of the lecithotrophic lizard Niveoscincus coventryi. As uterine angiogenesis and epithelial cell morphology are thought to be under progesterone control, we studied the effect of a progesterone receptor antagonist (mifepristone) on uterine and chorioallantoic microvasculature and features of the uterine epithelial surfaces. Although intussuceptive angiogenesis was observed in both, uterine and chorioallantoic, vascular beds during gestation, the only significant increases were in the diameters of the uterine vessels. An ellipsoid vessel-dense area grows in the mesometrial hemisphere of the developing conceptus, which parallels the expansion of the allantois to form the chorioallantoic placenta. Uterine surface topography changed during gestation. In particular, uterine blood vessels bulge over the luminal surface to form marked ridges on the uterine embryonic hemisphere, especially during the last stage of pregnancy, and ciliated cells are maintained in the embryonic and abembryonic hemispheres but disappear in both the mesometrial and antimesometrial poles. This distinct regionalization of uterine ridges and ciliated cells in the uterine surface and in the shape of the epithelial component of the chorion might be related to the function of both chorioallantoic and yolk sac placentae during gestation. There was no significant difference between females treated with or without mifepristone, which may be related to the partial function of mifepristone as a progestin antagonist and/or with the function and time of action of progesterone in the uterus during gestation in N. coventryi. Differences in the pattern of angiogenesis and uterine surface morphology during gestation among squamates may be related to the functional diversity of the uterine component of the different placentae and probably reflect its diverse evolutionary history. J. Morphol., 2011. © 2011 Wiley Periodicals, Inc.
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angiogenesis of the uterus and Chorioallantois in the eastern water skink eulamprus quoyii
The Journal of Experimental Biology, 2010Co-Authors: Bridget F Murphy, Scott L Parker, Christopher R Murphy, Michael B ThompsonAbstract:SUMMARY We have discovered a modification of the uterus that appears to facilitate maternal-fetal communication during pregnancy in the scincid lizard Eulamprus quoyii. A vessel-dense elliptical area (VDE) on the mesometrial side of the uterus expands as the embryo grows, providing a large vascular area for physiological exchange between mother and embryo. The VDE is already developed in females with newly ovulated eggs, and is situated directly adjacent to the Chorioallantois of the embryo when it develops. It is likely that signals from the early developing embryo determine the position of the VDE, as the VDE is off-centre in cases where the embryo sits obliquely in the uterus. The VDE is not a modification of the uterus over the entire chorioallantoic placenta, as the VDE is smaller than the Chorioallantois after embryonic stage 33, but expansion of the VDE and growth of the Chorioallantois during pregnancy are strongly correlated. The expansion of the VDE is also strongly correlated with embryonic growth and increasing embryonic oxygen demand (). We propose that angiogenic stimuli are exchanged between the VDE and the Chorioallantois in E. quoyii, allowing the simultaneous growth of both tissues.
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055 even reptiles do it the structure and function of placentae in viviparous lizards
Reproduction Fertility and Development, 2010Co-Authors: Michael B ThompsonAbstract:Live birth (viviparity) has evolved independently more than 100 times in squamate reptiles (lizards and snakes). Most viviparous lineages are characterised by a simple placenta with squamous epithelia on both maternal and embryonic sides, and the remnants of an eggshell persist for some, if not all, of pregnancy. The embryos are predominantly lecithotrophic, with maternal-embryonic exchanges being limited mostly to inorganic ions, water and respiratory gases. Nevertheless, there is differentiation of a chorioallantoic placenta and a yolk sac (omphalo) placenta in all species. Complex placentae have evolved in the Squamata in only four or five lineages, all in the lizard family Scincidae (skinks). In species with complex placentation, the uterus differentiates to allow different functions in association with each embryonic membrane type. Species with complex placentae are characterised by a hypertrophy of maternal and embryonic cells, elaboration of the maternal surface, a reduction in yolk with a concomitant increase in placentotrophy and, in some, regional differentiation of the choriallantois into a placentome and a paraplacentome. Both the placentome and omphaloplacenta are organs of embryonic nutrition, but they transport nutrients by different mechanisms; the paraplacentome is a specialised gas exchange organ. The most placentotrophic species are in the South American genus Mabuya, where the females produce micro-lecithal eggs and the placenta is more complex than in any mammal, with four specialised structures for nutrient exchange and one for gas exchange. The number of independent origins of viviparity and the range of placental complexities exhibited by skinks enables us to infer the evolutionary trajectories that have resulted in microlecithal eggs and an almost complete reliance of placentotrophy from oviparous ancestors.