28
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Choroid Sprouting Assay: An Ex Vivo Model of Microvascular Angiogenesis

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Angiogenesis of the microvasculature is central to the etiology of many diseases including proliferative retinopathy, age-related macular degeneration and cancer. A mouse model of microvascular angiogenesis would be very valuable and enable access to a wide range of genetically manipulated tissues that closely approximate small blood vessel growth in vivo. Vascular endothelial cells cultured in vitro are widely used, however, isolating pure vascular murine endothelial cells is technically challenging. A microvascular mouse explant model that is robust, quantitative and can be reproduced without difficulty would overcome these limitations. Here we characterized and optimized for reproducibility an organotypic microvascular angiogenesis mouse and rat model from the choroid, a microvascular bed in the posterior of eye. The choroidal tissues from C57BL/6J and 129S6/SvEvTac mice and Sprague Dawley rats were isolated and incubated in Matrigel. Vascular sprouting was comparable between choroid samples obtained from different animals of the same genetic background. The sprouting area, normalized to controls, was highly reproducible between independent experiments. We developed a semi-automated macro in ImageJ software to allow for more efficient quantification of sprouting area. Isolated choroid explants responded to manipulation of the external environment while maintaining the local interactions of endothelial cells with neighboring cells, including pericytes and macrophages as evidenced by immunohistochemistry and fluorescence-activated cell sorting (FACS) analysis. This reproducible ex vivo angiogenesis assay can be used to evaluate angiogenic potential of pharmacologic compounds on microvessels and can take advantage of genetically manipulated mouse tissue for microvascular disease research.

          Related collections

          Most cited references32

          • Record: found
          • Abstract: found
          • Article: not found

          Prevalence of age-related macular degeneration in the United States.

          To estimate the prevalence and distribution of age-related macular degeneration (AMD) in the United States by age, race/ethnicity, and gender. Summary prevalence estimates of drusen 125 microm or larger, neovascular AMD, and geographic atrophy were prepared separately for black and white persons in 5-year age intervals starting at 40 years. The estimated rates were based on a meta-analysis of recent population-based studies in the United States, Australia, and Europe. These rates were applied to 2000 US Census data and to projected US population figures for 2020 to estimate the number of the US population with drusen and AMD. The overall prevalence of neovascular AMD and/or geographic atrophy in the US population 40 years and older is estimated to be 1.47% (95% confidence interval, 1.38%-1.55%), with 1.75 million citizens having AMD. The prevalence of AMD increased dramatically with age, with more than 15% of the white women older than 80 years having neovascular AMD and/or geographic atrophy. More than 7 million individuals had drusen measuring 125 microm or larger and were, therefore, at substantial risk of developing AMD. Owing to the rapidly aging population, the number of persons having AMD will increase by 50% to 2.95 million in 2020. Age-related macular degeneration was far more prevalent among white than among black persons. Age-related macular degeneration affects more than 1.75 million individuals in the United States. Owing to the rapid aging of the US population, this number will increase to almost 3 million by 2020.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Phenotypic heterogeneity of the endothelium: II. Representative vascular beds.

            Endothelial cells, which form the inner cellular lining of blood vessels and lymphatics, display remarkable heterogeneity in structure and function. This is the second of a 2-part review on the phenotypic heterogeneity of blood vessel endothelial cells. The first part discusses the scope, the underlying mechanisms, and the diagnostic and therapeutic implications of phenotypic heterogeneity. Here, these principles are applied to an understanding of organ-specific phenotypes in representative vascular beds including arteries and veins, heart, lung, liver, and kidney. The goal is to underscore the importance of site-specific properties of the endothelium in mediating homeostasis and focal vascular pathology, while at the same time emphasizing the value of approaching the endothelium as an integrated system.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Targeting the tumour vasculature: insights from physiological angiogenesis.

              The cardiovascular system ensures the delivery of nutrients, oxygen, and blood and immune cells to all organs and tissues: it is also responsible for the removal of waste metabolites. The vascular system develops and matures through two tightly regulated processes: vasculogenesis and angiogenesis. Angiogenesis is active only under specific physiological conditions in healthy adults but the vasculature can be aberrantly activated to generate new blood vessels during pathological conditions such as cancer and chronic inflammation. In this Opinion article we discuss the parallels and differences in the angiogenic process under either a physiological or a pathological state, especially tumorigenesis.
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2013
                26 July 2013
                : 8
                : 7
                : e69552
                Affiliations
                [1 ]Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America
                [2 ]Department of Ophthalmology, Research Centers of Hôpital Maisonneuve-Rosemont, University of Montreal, Montreal, Quebec, Canada
                [3 ]Departments of Pediatrics Ophthalmology and Pharmacology, Research Centers of CHU Sainte-Justine, Montreal, Quebec, Canada
                Institut de la Vision, France
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: ZS MF JC JSJ LS. Performed the experiments: ZS MF CH ZC DP LE AJ. Analyzed the data: ZS MF CH ZC DP LE AJ PS LS. Contributed reagents/materials/analysis tools: HT FD SC. Wrote the paper: ZS JC PS JSJ LS.

                Article
                PONE-D-13-18260
                10.1371/journal.pone.0069552
                3724908
                23922736
                94a1973a-63dd-4862-8ce8-72f3b8bafbb9
                Copyright @ 2013

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 2 May 2013
                : 11 June 2013
                Page count
                Pages: 11
                Funding
                This work is supported by funding from National Institutes of Health (NIH) (EY017017-04S1, EY017017-05), V. Kann Rasmussen Foundation, Boston Children's Hospital Mental Retardation and Developmental Disabilities Research Center PO1HD18655, Research to Prevent Blindness Senior Investigator Award, Alcon Research Institute Award and Lowy Medical Foundation (LEHS). ZS is supported by Canadian Institute of Health Research (CIHR) System Biology Training Scholarship and Travel Grant. FD is supported by CIHR Banting & Best PhD Scholarship and Health Professional PhD Scholarship from the Fonds de la Recherche en Santé du Québec (FRSQ). JC is supported by Boston Children's Hospital Ophthalmology Foundation, Charles H. Hood Foundation Child Health Research Award, Blind Children's Center, and BrightFocus Foundation. PS holds a Canada Research Chair tier II. SC holds a Canada Research chair tier I (Vision science) and the Leopoldine Wolfe Chair in translational research in macular degeneration (U Montreal). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Model Organisms
                Animal Models
                Mouse
                Rat
                Molecular Cell Biology
                Cellular Types
                Endothelial Cells
                Cell Growth
                Medicine
                Cardiovascular
                Vascular Biology
                Ophthalmology
                Retinal Disorders

                Uncategorized
                Uncategorized

                Comments

                Comment on this article