scholarly journals Selective Binding of Lectins to Embryonic Chicken Vasculature

2003 ◽  
Vol 51 (5) ◽  
pp. 597-604 ◽  
Author(s):  
Shahla M. Jilani ◽  
Thomas J. Murphy ◽  
Shelley N.M. Thai ◽  
Anne Eichmann ◽  
Jackelyn A. Alva ◽  
...  
Keyword(s):  
Lens Culinaris ◽  
Developmental Stages ◽  
Chorioallantoic Membrane ◽  
Experimental System ◽  
In Ovo ◽  
Additional Advantage ◽  
Chicken Embryos ◽  
Lens Culinaris Agglutinin ◽  
Embryonic Chicken

Chicken embryos are an excellent model system for studies related to vascular morphogenesis. Development in ovo allows manipulations otherwise difficult in mammals, and the use of chicken-quail chimeras offers an additional advantage to this experimental system. Furthermore, the chicken chorioallantoic membrane has been extensively used for in vivo assays of angiogenesis. Surprisingly, few markers are available for a comprehensive visualization of the vasculature. Here we report the use of lectins for identification of embryonic chicken blood vessels. Nine lectins were evaluated using intravascular perfusion and directly on sections. Our results indicate that Lens culinaris agglutinin, concanavalin A, and wheat germ agglutinin can be used effectively for visualization of vessels of early chicken embryos (E2.5-E4). At later developmental stages, Lens culinaris agglutinin is a better choice because it displays equal affinity for the endothelia of arteries, veins, and capillaries. The findings presented here expand our understanding of lectin specificity in the endothelium of avian species and provide information as to the use of these reagents to obtain comprehensive labeling of the embryonic and chorioallantoic membrane vasculature.

scholarly journals Improved Method forEx Ovo-Cultivation of Developing Chicken Embryos for Human Stem Cell Xenografts

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Timo Schomann ◽  
Firas Qunneis ◽  
Darius Widera ◽  
Christian Kaltschmidt ◽  
Barbara Kaltschmidt
Keyword(s):  
Stem Cells ◽  
Chicken Embryo ◽  
Adult Stem Cells ◽  
Developmental Stages ◽  
Human Stem Cells ◽  
In Ovo ◽  
Differentiation Potential ◽  
Chicken Embryos

The characterization of human stem cells for the usability in regenerative medicine is particularly based on investigations regarding their differentiation potentialin vivo. In this regard, the chicken embryo model represents an ideal model organism. However, the access to the chicken embryo is only achievable by windowing the eggshell resulting in limited visibility and accessibility in subsequent experiments. On the contrary,ex ovo-culture systems avoid such negative side effects. Here, we present an improvedex ovo-cultivation method enabling the embryos to survive 13 daysin vitro. Optimized cultivation of chicken embryos resulted in a normal development regarding their size and weight. Ourex ovo-approach closely resembles the development of chicken embryosin ovo, as demonstrated by properly developed nervous system, bones, and cartilage at expected time points. Finally, we investigated the usability of our method for trans-species transplantation of adult stem cells by injecting human neural crest-derived stem cells into late Hamburger and Hamilton stages (HH26–HH28/E5—E6) ofex ovo-incubated embryos. We demonstrated the integration of human cells allowing experimentally easy investigation of the differentiation potential in the proper developmental context. Taken together, thisex ovo-method supports the prolonged cultivation of properly developing chicken embryos enabling integration studies of xenografted mammalian stem cells at late developmental stages.

scholarly journals The Role of Pre-Clinical 3-Dimensional Models of Osteosarcoma

2020 ◽  
Vol 21 (15) ◽  
pp. 5499
Author(s):  
Hannah L. Smith ◽  
Stephen A. Beers ◽  
Juliet C. Gray ◽  
Janos M. Kanczler
Keyword(s):  
Three Dimensional ◽  
Chorioallantoic Membrane ◽  
3D Models ◽  
In Ovo ◽  
Future Directions ◽  
3 Dimensional ◽  
In Vivo Animal Models

Treatment for osteosarcoma (OS) has been largely unchanged for several decades, with typical therapies being a mixture of chemotherapy and surgery. Although therapeutic targets and products against cancer are being continually developed, only a limited number have proved therapeutically active in OS. Thus, the understanding of the OS microenvironment and its interactions are becoming more important in developing new therapies. Three-dimensional (3D) models are important tools in increasing our understanding of complex mechanisms and interactions, such as in OS. In this review, in vivo animal models, in vitro 3D models and in ovo chorioallantoic membrane (CAM) models, are evaluated and discussed as to their contribution in understanding the progressive nature of OS, and cancer research. We aim to provide insight and prospective future directions into the potential translation of 3D models in OS.

Nerve-dependent accumulation of myosin light chain 3 in developing limb musculature

Development ◽  
1987 ◽  
Vol 101 (4) ◽  
pp. 673-684
Author(s):  
P.A. Merrifield ◽  
I.R. Konigsberg
Keyword(s):  
Neural Tube ◽  
Light Chain ◽  
Limb Development ◽  
Myosin Light Chain ◽  
Chorioallantoic Membrane ◽  
In Ovo ◽  
Fast Myosin ◽  
Limb Musculature

Myosin alkali light chain accumulation in developing quail limb musculature has been analysed on immunoblots using a monoclonal antibody which recognizes an epitope common to fast myosin light chain 1 (MLC1f) and fast myosin light chain 3 (MLC3f). The limb muscle of early embryos (i.e. up to day 10 in ovo) has a MLC profile similar to that observed in myotubes cultured in vitro; although MLC1f is abundant, MLC3f cannot be detected. MLC3f is first detected in 11-day embryos. To determine whether this alteration in MLC3f accumulation is nerve or hormone dependent, limb buds with and without neural tube were cultured as grafts on the chorioallantoic membrane of chick hosts. Although differentiated muscle develops in both aneural and innervated grafts, innervated grafts contain approximately three times as much myosin as aneural grafts. More significantly, although aneural grafts reproducibly accumulate normal levels of MLC1f, they fail to accumulate detectable levels of MLC3f. In contrast, innervated grafts accumulate both MLC1f and MLC3f, suggesting that the presence of neural tube in the graft promotes the maturation, as well as the growth, of muscle tissue. This is the first positive demonstration that innervation is necessary for the accumulation of MLC3f that occurs during normal limb development in vivo.

scholarly journals Extending the Applicability of In Ovo and Ex Ovo Chicken Chorioallantoic Membrane Assays to Study Cytostatic Activity in Neuroblastoma Cells

2021 ◽  
Vol 11 ◽  
Author(s):  
Miguel Angel Merlos Rodrigo ◽  
Berta Casar ◽  
Hana Michalkova ◽  
Ana Maria Jimenez Jimenez ◽  
Zbynek Heger ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Neuroblastoma Cells ◽  
Chorioallantoic Membrane ◽  
Stage Iv ◽  
Metastatic Potential ◽  
Significant Inhibition ◽  
Cam Assay ◽  
Anticancer Activities ◽  
In Ovo

PurposeThe chick chorioallantoic membrane (CAM) assay can provide an alternative versatile, cost-effective, and ethically less controversial in vivo model for reliable screening of drugs. In the presented work, we demonstrate that CAM assay (in ovo and ex ovo) can be simply employed to delineate the effects of cisplatin (CDDP) and ellipticine (Elli) on neuroblastoma (Nbl) cells in terms of their growth and metastatic potential.MethodsThe Nbl UKF-NB-4 cell line was established from recurrent bone marrow metastases of high-risk Nbl (stage IV, MYCN amplification, 7q21 gain). Ex ovo and in ovo CAM assays were optimized to evaluate the antimetastatic activity of CDDP and Elli. Immunohistochemistry, qRT-PCR, and DNA isolation were performed.ResultsEx ovo CAM assay was employed to study whether CDDP and Elli exhibit any inhibitory effects on growth of Nbl xenograft in ex ovo CAM assay. Under the optimal conditions, Elli and CDDP exhibited significant inhibition of the size of the primary tumor. To study the efficiency of CDDP and Elli to inhibit primary Nbl tumor growth, intravasation, and extravasation in the organs, we adapted the in ovo CAM assay protocol. In in ovo CAM assay, both studied compounds (CDDP and Elli) exhibited significant (p < 0.001) inhibitory activity against extravasation to all investigated organs including distal CAM.ConclusionsTaken together, CAM assay could be a helpful and highly efficient in vivo approach for high-throughput screening of libraries of compounds with expected anticancer activities.

scholarly journals The Chorioallantoic Membrane Assay in Nanotoxicological Research—An Alternative for In Vivo Experimentation

Nanomaterials ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2328
Author(s):  
Christoph R. Buhr ◽  
Nadine Wiesmann ◽  
Rachel C. Tanner ◽  
Jürgen Brieger ◽  
Jonas Eckrich
Keyword(s):  
Imaging Techniques ◽  
Chorioallantoic Membrane ◽  
Drug Carriers ◽  
Human Organism ◽  
Cam Assay ◽  
In Ovo ◽  
In Vivo Models ◽  
Toxicological Profile ◽  
Application Fields

Nanomaterials unveil many applicational possibilities for technical and medical purposes, which range from imaging techniques to the use as drug carriers. Prior to any human application, analysis of undesired effects and characterization of their toxicological profile is mandatory. To address this topic, animal models, and rodent models in particular, are most frequently used. However, as the reproducibility and transferability to the human organism of animal experimental data is increasingly questioned and the awareness of animal welfare in society increases at the same time, methodological alternatives are urgently required. The chorioallantoic membrane (CAM) assay is an increasingly popular in ovo experimental organism suitable for replacement of rodent experimentation. In this review, we outline several application fields for the CAM assay in the field of nanotoxicology. Furthermore, analytical methods applicable with this model were evaluated in detail. We further discuss ethical, financial, and bureaucratic aspects and benchmark the assay with other established in vivo models such as rodents.

scholarly journals Functional involvement of carbonic anhydrase in calcium transport of the chick chorioallantoic membrane

1978 ◽  
Vol 176 (1) ◽  
pp. 67-74 ◽  
Author(s):  
R S Tuan ◽  
J Zrike
Keyword(s):  
Carbonic Anhydrase ◽  
Chorioallantoic Membrane ◽  
Carbonic Anhydrase Activity ◽  
Calcium Deposition ◽  
Dependent Manner ◽  
Transport Activity ◽  
In Ovo ◽  
Chick Chorioallantoic Membrane ◽  
Age Dependent

Carbonic anhydrase activity was demonstrated in the chick-embryonic chorioallantoic membrane and was correlated with the Ca2+-transport activity of the membrane. It is inhibited by sulphonamides and is expressed in the chorioallantoic membrane in an age-dependent fashion during embryonic development. Ca2+ uptake by the chorioallantoic membrane in vivo also increases in a similar age-dependent manner. The temporal increase in these activities is coincident with calcium deposition in the embryonic skeleton. Incubation of the chorioallantoic membrane in ovo with sulphonamides specifically inhibits both the carbonic anhydrase and the Ca2+ uptake activities of the membrane in vivo. Enzyme histochemistry revealed the carbonic anhydrase activity is localized in the Ca2+-transporting ectodermal cells of the chorioallantoic membrane. These results, taken together, indicate that carbonic anhydrase may be functionally important in the Ca2+-transport activity of the chorioallantoic membrane.

Calcium-transport function of the chick embryonic chorioallantoic membrane. I. In vivo and in vitro characterization

1986 ◽  
Vol 82 (1) ◽  
pp. 73-84
Author(s):  
R.S. Tuan ◽  
M.J. Carson ◽  
J.A. Jozefiak ◽  
K.A. Knowles ◽  
B.A. Shotwell
Keyword(s):  
Calcium Transport ◽  
Calcium Uptake ◽  
Chorioallantoic Membrane ◽  
Transport Function ◽  
In Ovo ◽  
In Vitro Characterization ◽  
Microsomal Membranes

During chick embryonic development, the chorioallantoic membrane (CAM) is responsible for the mobilization of shell calcium into the embryonic circulation. The calcium-transport function of the CAM was studied here by measuring CAM calcium uptake in vivo and in vitro. The in vivo technique involved the use of an uptake chamber constructed on top of the CAM in situ. The in vitro methods included two systems: CAM tissue disks and cell-free microsomal membranes isolated from the CAM. Analyses using these three assays show that calcium uptake by the CAM exhibited characteristics indicative of active transport, such as temperature dependence, saturability, energetic requirement and ion specificity. The data also show that calcium-uptake activities of the CAM increase as a function of embryonic age in a manner coincident with the increased accumulation of calcium by the developing embryo in ovo.

scholarly journals Angiogenic Effect of Leptin in the Quail Chorioallantoic Membrane

2010 ◽  
Vol 79 (1) ◽  
pp. 13-17 ◽  
Author(s):  
Pavel Výboh ◽  
Michal Zeman ◽  
Boris Bilčík ◽  
Božena Šárniková ◽  
Ľubor Košťál
Keyword(s):  
Embryonic Development ◽  
Growth And Development ◽  
Chorioallantoic Membrane ◽  
Nutrient Utilization ◽  
Avian Embryo ◽  
In Ovo ◽  
Angiogenic Effect ◽  
Vessel Development ◽  
Stimulation Of

Leptin, the product ofobgene, beside its key role in the control of body weight and food consumption, can be involved in the control of embryonic development. Leptin administrationin ovoaccelerated the embryonic and post-embryonic development in Japanese quail. Although the mechanisms of leptin effects on growth and development acceleration are not clear, stimulation of angiogenesis represents one of plausible explanations. Therefore, the aim of the present study was to investigate the pro-angiogenic effect of leptinin vivoin the quail chorioallantoic membrane (CAM). The recombinant murine leptin (10, 100, and 1000 ng) was applied eitherex ovoon the CAM surface ofex ovoincubated embryos at embryonic day 7 (ED7) orin ovointo the egg albumen at ED5. Changes in blood vessels were quantified by the fractal analysis providing the fractal dimension (Df) estimate. Leptin administeredin ovowas more efficient in stimulation of angiogenesis than theex ovotreatment, since 10 ng dose elicited significantly higher (P< 0.001) stimulation of vessel development of the CAM under the air cell than it did afterex ovocultivation. Our study confirmed that exogenously applied leptin was able to stimulate angiogenesis in CAM. Leptin-mediated stimulation of angiogenesis may improve nutrient utilization from the yolk and explain at least partially the accelerating effect of leptin on avian embryo growth and development.

scholarly journals Monitoring of tumor growth and vascularization with repetitive ultrasonography in the chicken chorioallantoic-membrane-assay

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jonas Eckrich ◽  
Philipp Kugler ◽  
Christoph Raphael Buhr ◽  
Benjamin Philipp Ernst ◽  
Simone Mendler ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Tumor Imaging ◽  
Imaging Techniques ◽  
Chorioallantoic Membrane ◽  
Cam Assay ◽  
In Ovo ◽  
Tumor Research ◽  
Xenograft Models ◽  
Cost Efficient

Abstract The chorioallantoic-membrane (CAM)-assay is an established model for in vivo tumor research. Contrary to rodent-xenograft-models, the CAM-assay does not require breeding of immunodeficient strains due to native immunodeficiency. This allows xenografts to grow on the non-innervated CAM without pain or impairment for the embryo. Considering multidirectional tumor growth, limited monitoring capability of tumor size is the main methodological limitation of the CAM-assay for tumor research. Enclosure of the tumor by the radiopaque eggshell and the small structural size only allows monitoring from above and challenges established imaging techniques. We report the eligibility of ultrasonography for repetitive visualization of tumor growth and vascularization in the CAM-assay. After tumor ingrowth, ultrasonography was repetitively performed in ovo using a commercial ultrasonographic scanner. Finally, the tumor was excised and histologically analyzed. Tumor growth and angiogenesis were successfully monitored and findings in ultrasonographic imaging significantly correlated with results obtained in histological analysis. Ultrasonography is cost efficient and widely available. Tumor imaging in ovo enables the longitudinal monitoring of tumoral development, yet allowing high quantitative output due to the CAM-assays simple and cheap methodology. Thus, this methodological novelty improves reproducibility in the field of in vivo tumor experimentation emphasizing the CAM-assay as an alternative to rodent-xenograft-models.

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