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• W1993235364 abstract "HomeCirculation ResearchVol. 116, No. 11State-of-the-Art Methods for Evaluation of Angiogenesis and Tissue Vascularization Free AccessResearch ArticlePDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessResearch ArticlePDF/EPUBState-of-the-Art Methods for Evaluation of Angiogenesis and Tissue VascularizationA Scientific Statement From the American Heart Association Michael Simons, MD, Kari Alitalo, MD, PhD, Brian H. Annex, MD, Hellmut G. Augustin, MD, Craig Beam, CRE, Bradford C. Berk, MD, PhD, FAHA, Tatiana Byzova, PhD, Peter Carmeliet, MD, PhD, William Chilian, PhD, FAHA, John P. Cooke, MD, George E. Davis, MD, PhD, Anne Eichmann, PhD, M. Luisa Iruela-Arispe, PhD, Eli Keshet, PhD, Albert J. Sinusas, MD, FAHA, Christiana Ruhrberg, PhD, Y. Joseph Woo, MD, FAHA and Stefanie Dimmeler, MDon behalf of the American Heart Association Council on Basic Cardiovascular Sciences and Council on Cardiovascular Surgery and Anesthesia Michael SimonsMichael Simons Search for more papers by this author , Kari AlitaloKari Alitalo Search for more papers by this author , Brian H. AnnexBrian H. Annex Search for more papers by this author , Hellmut G. AugustinHellmut G. Augustin Search for more papers by this author , Craig BeamCraig Beam Search for more papers by this author , Bradford C. BerkBradford C. Berk Search for more papers by this author , Tatiana ByzovaTatiana Byzova Search for more papers by this author , Peter CarmelietPeter Carmeliet Search for more papers by this author , William ChilianWilliam Chilian Search for more papers by this author , John P. CookeJohn P. Cooke Search for more papers by this author , George E. DavisGeorge E. Davis Search for more papers by this author , Anne EichmannAnne Eichmann Search for more papers by this author , M. Luisa Iruela-ArispeM. Luisa Iruela-Arispe Search for more papers by this author , Eli KeshetEli Keshet Search for more papers by this author , Albert J. SinusasAlbert J. Sinusas Search for more papers by this author , Christiana RuhrbergChristiana Ruhrberg Search for more papers by this author , Y. Joseph WooY. Joseph Woo Search for more papers by this author and Stefanie DimmelerStefanie Dimmeler Search for more papers by this author and on behalf of the American Heart Association Council on Basic Cardiovascular Sciences and Council on Cardiovascular Surgery and Anesthesia Originally published30 Apr 2015https://doi.org/10.1161/RES.0000000000000054Circulation Research. 2015;116:e99–e132Other version(s) of this articleYou are viewing the most recent version of this article. Previous versions: January 1, 2015: Previous Version 1 IntroductionVascular dysfunction is causally contributing to many diseases, including but not limited to cardiovascular disease, which is still the leading cause of death in the Western world. The endothelium that lines the inner wall of the blood vessels plays a critical role in the pathobiology of these illnesses. Particularly after ischemia or injury, the growth of new blood vessels, driven by endothelial expansion, is essential to maintain oxygen supply to the ischemic or injured tissue. Recent studies additionally suggest that the endothelium acts as a paracrine source for signals that determine tissue regeneration versus fibrosis after injury. Excessive vascularization, however, might also be unwanted, as in the case of cancer, neovascular eye diseases including diabetic retinopathy, atheroma growth, or the expansion of vasa vasorum, which leads to adverse vessel wall remodeling.Neovascularization is a tightly regulated and essential process that results in the formation of new blood vessels. Specific types of neovascularization include angiogenesis, the formation of new capillaries from existing capillaries, and arteriogenesis, the formation of new arteries from preexisting collaterals or de novo. Although endothelial cells (ECs) certainly are essential for both processes, the formation of functionally active vessels requires a complex molecular cross-talk of ECs with perivascular cells such as pericytes, smooth muscle cells, and macrophages. Simple in vitro models are best suited to examine specific aspects of particular processes involved in angiogenesis such as the biochemical interactions that regulate EC proliferation, motility, and apoptosis or lumen formation. However, in vivo models are required to understand the complex cellular interactions that enable the generation of functionally active and hierarchical blood vessel networks capable of providing an appropriate blood supply and paracrine stimuli to organs. In addition, the development of therapeutic strategies to either promote or inhibit vessel growth depends on reproducible measures and end points in experimental models that are relevant for the treatment of human diseases. Accordingly, the selection of an appropriate experimental model is critical to study specific aspects of the molecular and cellular mechanisms that are of physiological or pathological relevance. This scientific statement summarizes in vitro assays and in vivo models suitable for gaining insights into the basic mechanisms of vessel growth and provides an overview of models that are particularly useful for clinical translation.1. In Vitro AssaysConsiderable progress has been made over the past 20 years in the development of in vitro strategies to investigate the complex processes of vascular morphogenesis and maturation events in 3-dimensional (3D) extracellular matrixes1,2 (Table 1). Most studies have used primary human ECs, in particular human umbilical vein ECs,3–5 because they are easily available. Other studies have used the aortic ring assay from rat or mouse to study these processes.6 Two major extracellular matrix environments that work well in these assays are 3D type I collagen and fibrin matrixes. Both types of matrixes are highly effective in inducing EC tube formation and sprouting and allow EC-pericyte interactions in models of vessel maturation, which occur in a 3D matrix environment.4,7Table 1. Summary of Most Commonly Used In Vitro AssaysProCon3D collagen orfibrin EC lumen formation and sprouting assaysVery reproducibleAllows the study of EC tube formation and sprouting in physiologically relevant 3D matrix environmentsCan be reproducibly performed under serum-free conditions with defined recombinant growth factorsCan be performed with various human ECs (eg, HUVECs) that can be genetically manipulatedEC tubes that are formed can be subjected to flow forcesEC only conditions without mural cellsThus far, isolated rodent ECs have not worked well in these assay modelsVariable growth conditions for ECs may affect results with these modelsEC-pericyte coculture assays in 3D collagen or fibrin matrixesAllows the study of EC tubulogenesis and sprouting and the contribution of mural cells to these eventsAllows the study of ECM remodeling, including basement membrane matrix assembly around developing EC tubesCan be performed under serum-free defined conditions and functionally define ECs vs pericytesCan be difficult to establish; functionality depends on appropriate conditionsPotential variability concerning the sources of pericytes used for the assay systemsEC spheroid assayVery reproducibleAngiogenic sprouting can be investigatedNo mural cell contributionAortic ring assayVery reproducibleAllows the study of the angiogenic effects of genes in genetic models ex vivoVariability requires sufficiently high numbersCellular contributions (eg, ECs, various mural cells, macrophages) may varyMatrigel assayVery reproducibleCan provide insight into EC survival and signaling activityIs not a specific angiogenesis assay (other cells can also form cordlike networks)No tubulogenesis3D indicates 3-dimensional; EC, endothelial cell; ECM, extracellular matrix; and HUVEC, human umbilical vein endothelial cell.Many studies have also been performed by seeding ECs 2-dimensionally on the surface of Matrigel, a laminin-rich matrix,8 which enables ECs to rearrange into cordlike, lumenless structures.9 However, there is actually little evidence that this assay system provides a suitable in vitro representation of the process of vascular morphogenesis. In fact, many cell types, including vascular smooth muscle cells, fibroblasts, and tumor cells, form similar cordlike structures on Matrigel surfaces, suggesting that this alignment assay may reflect a competitive cell-cell versus cell-matrix adhesion assay.9 Laminin-rich matrixes have been shown to inhibit tubulogenesis through integrin-laminin interactions.10 Together, 3D collagen and fibrin models have been demonstrated to be superior for investigations of vascular tubulogenesis, sprouting, and EC-pericyte tube coassembly and maturation events, including vascular basement membrane matrix formation.1.1. EC Tubulogenesis ModelsA fundamental property of ECs is their ability to form tubes with lumen structures in 3D matrixes (ie, a property of arterial, capillary, venous, and lymphatic ECs).1 Any cell type that is reported to be an EC (including cells derived from induced pluripotent stem cells or other stem/progenitor cells that have differentiated toward the EC lineage) should be able to form tubes in a 3D matrix environment in vitro. We propose that without such functional characterization, a cell type should not be classified as an EC even if it expresses EC markers.Several model systems have been established over many years to elucidate fundamental mechanisms underlying how ECs assemble into tube structures with defined lumens in 3D collagen or fibrin matrixes.3,4 In the most basic setup, ECs are seeded into a 3D matrix and align over time to form lumenized tubes. Tube formation has been demonstrated by cross sections of cultures and with the use of techniques such as thin sectioning of plastic embedded gels, confocal microscopy, and transmission electron microscopy. Importantly, one consequence of EC tubulogenesis is the creation of physical tunnel spaces in the extracellular matrix, called vascular guidance tunnels, that are generated through highly localized MT1-matrix metalloproteinase–dependent matrix degradation.11,12 These tunnel spaces represent matrix conduits for EC motility, tube remodeling, and recruitment of mural cells to the abluminal surface of developing vessels to enable processes such as vessel remodeling and vascular basement membrane matrix assembly.13In vitro models have been used as a primary discovery tool to identify critical regulators of the EC lumen and tube formation pathway, which were later confirmed with in vivo models. They have also have been extensively used to confirm in vivo findings that have been obtained through mouse knockout or zebrafish morpholino experiments. One point that should be emphasized is that in vitro morphogenesis models with isolated ECs have used human ECs; thus, if differences arise with an in vivo model system, it is always possible that this discordance may reflect a species difference in the molecular requirements for vascular morphogenic events.To date, it has been very difficult to successfully use isolated primary mouse or rat ECs in tube morphogenic assays in 3D collagen or fibrin matrixes. The reason for this discrepancy between human and rodent ECs is unclear. However, primary human ECs work extremely well in vascular tube morphogenesis and sprouting assays in 3D matrixes.As a general rule, in vitro vascular morphogenesis assays should be performed under highly defined conditions. Ideally, the assay systems should be performed in serum-free defined media in 3D matrixes so that the additives are known, and this has been accomplished with the use of a number of in vitro models using human ECs and the aortic ring assay. In this manner, the actual growth factor, lipid, hormone, and so on, requirements for the various steps and events during vascular morphogenesis can be accurately assessed.1.2. Sprouting ModelsIn contrast to 3D alignment assays, the process of sprouting angiogenesis can be studied in vitro by allowing ECs to form sprouting and lumen-forming capillary-like structures by seeding them from focal starting points into a collagen or fibrin matrix. A common strategy to focally seed ECs into a matrix is to coat beads with ECs and then embed these EC-coated beads into a 3D matrix.14 Alternatively, ECs can be allowed to form 3D spheroidal aggregates that can similarly be delivered as focal starting points into a 3D matrix.5 Spheroidal aggregates of human ECs have also been transplanted in a 3D matrix into immunocompromised mice to generate a 3D capillary network that anastomoses with the mouse vasculature and is perfused by mouse blood.15,16 Sprouting angiogenesis can be demonstrated and qualitatively and quantitatively analyzed effectively in vitro and in vivo in these assays.Sprouting angiogenesis has also been studied by allowing ECs seeded as monolayers on top of a matrix to invade into the matrix, which allows them to form lumenized structures.17,18 Such 2-dimensional/3D hybrid assays have been established as 3D sprouting angiogenesis assays. They can be considered an appropriate representation of 3D sprouting angiogenesis. However, the proteolytic balance between matrix-degrading and matrix-invading forces needs to carefully controlled.The aortic ring assay system is another widely used 3D sprouting angiogenesis assay.6 This assay has some advantages in that animal tissues from normal versus genetically modified vessels can be directly assessed in an ex vivo assay. In turn, the multicellular nature of this assay can complicate the interpretation of results. The aortic ring assay has also been modified to study the sprouting of lymphatic ECs from explanted thoracic duct segments.19A murine allantois assay system that is useful for investigating early vasculogenic tube assembly has also been described, although no exogenous extracellular matrix is used in this approach.201.3. Microfluidic Flow ModelsA recent advance has been the development of microfluidic models of vascular morphogenesis whereby flow forces can be applied during the morphogenic event. To date, this has been performed primarily with sprouting models in which flow is applied to 1 monolayer surface and then the tube networks connect with 2 sides of the microfluidic device (containing flow channels).21 Directional flow ensues through the tubes that connect the flow channels. Other variations of such models have been to create tube channels artificially in 3D matrixes to enable user-defined geometries of the vascular network and then seed the channels with ECs for the creation of tube networks in the absence of EC-driven tubulogenesis and vascular guidance tunnel formation.22,23 Of note, these systems have so far not been tested by many laboratories, and the robustness and practicability at a larger scale remain to be established.1.4. EC-Pericyte Tube Maturation ModelsSeveral EC-pericyte coculture models in 3D matrixes have been established during the past 10 years. Such assays have highlighted the functional importance of both ECs and pericytes during capillary tube assembly in vitro.7,12,13,24 A major function of ECs is to establish networks of tubes that reside within vascular guidance tunnels and that induce the recruitment of pericytes into tunnel spaces along the abluminal tube surface.7 Both ECs and pericytes are highly motile during these events, and once pericytes reach the abluminal surface, they migrate along these surfaces in a polarized manner. Pericyte recruitment increases the length and decreases the diameter of EC tubes, which correlates with the deposition of vascular basement membrane matrix. Both cell types contribute basement membrane components, and both ECs and pericytes are necessary to generate basement membrane matrixes under defined serum-free conditions in 3D matrixes (demonstrated with transmission electron microscopy and immunofluorescence microscopy). Such systems enable a detailed analysis of EC tube maturation events and can assess the role of the key growth factors and signaling pathways that are necessary for these processes.1.5. SummaryOverall, in vitro models of vascular morphogenesis and maturation have contributed greatly to our understanding of these events, and sprouting angiogenesis can be well studied by the established highly reproducible assays. The use of 3D coculture assays additionally allows investigators to gain insights into vessel maturation processes and to study EC-pericyte interactions. The development of 3D coculture models that include flow would allow the study of microcirculatory networks and vessel maintenance at least for some days. Clearly, these models should be used in conjunction with in vivo approaches and hybrid strategies in which ECs are manipulated in vitro and then introduced in vivo to interface with a host vasculature. This is essential because even the best in vitro model will not fully recapitulate the complex process of vessel growth under physiological or pathophysiological conditions. Furthermore, in vitro models cannot assess the impact of circulating humoral and cellular factors that interact with the endothelium and that could have major impact, particularly under disease conditions such as diabetes mellitus, coronary artery disease, or heart failure, which systemically affect tissue homeostasis.2. Developmental Angiogenesis and ArteriogenesisIn all vertebrate species, blood vessels form during embryogenesis in successive steps called vasculogenesis and angiogenesis, which are then followed by remodeling and maturation of the initial vascular plexus into adult vasculature.25 The term vasculogenesis describes the de novo specification of endothelial precursor cells or angioblasts from the mesoderm. These newly formed cells coalesce into lumenized tubes that form the primary vascular plexus, which consists of a meshwork of homogeneously sized capillaries and the central axial vessels (ie, the dorsal aorta [DA] and cardinal veins). With the onset of heartbeat, this early vascular circuit expands and remodels, undergoing proliferation, sprouting, and pruning of preexisting vessels. This process of vascular remodeling transforms simple circulatory loops into a complex hierarchical network of branched endothelial tubes of varying diameter, length, and identity and depends on both genetically hard-wired events and hemodynamic forces (reviewed elsewhere26).A large number of signaling pathways regulate vascular development (reviewed by Potente et al27). Because a properly formed cardiovascular system is essential for sustaining embryonic development, mutations in these pathways frequently cause phenotypes that result in embryonic lethality. Global deletion in vascular endothelial growth factor (VEGF), Delta/Notch, integrin, transforming growth factor-ß, ephrin/Eph, and angiopoietin/Tie signaling pathways (among others) impairs vascular development, causing lethality in the mouse embryo between embryonic day (E) 8.0 and E12.5 attributable to failure to establish a functional circulatory network. These studies have been instrumental in our understanding of pathways required for vasculogenesis, endothelial tube assembly and lumen formation, angiogenesis, and arteriovenous differentiation. In the following sections, we present currently available models in mice and other species. Because vascular phenotypes are often complex and we still have an incomplete understanding of the normal sequence of vascular remodeling events, delineating the primary defect caused by a specific gene manipulation may require the combinatorial use of >1 model system.2.1. The Mouse Embryonic Hindbrain and Postnatal Retina as Model Systems to Study Developmental AngiogenesisThe developing mouse constitutes a model of choice to analyze vascular development and consequences of genetic manipulations on this process. Angioblasts in the mouse embryo first emerge from the mesoderm as VEGF receptor (VEGFR) 2+ cells around E7.0 and assemble into a simple circulatory loop consisting of a heart, DA, yolk sac plexus, and sinus venosus by E8.0.Several methodological improvements have allowed better understanding of early embryonic vascular development in the mouse. These include an expanded array of vascular markers and improved imaging techniques such as confocal imaging, optical projection tomography, and ultramicroscopy to visualize vascular networks in intact embryos with far greater degrees of resolution and 3D context compared with histological sections.28,29 In addition, the culture of allantois, an extraembryonic appendage rich in blood vessels that develops from E6 onward in the mouse, can be used for live imaging of vascular network formation in explanted wild-type and mutant tissue and allows additional manipulations such as phenotypic rescue experiments (reviewed by Arora and Papaioannou30).The most crucial technical innovation in recent years has been the generation of temporally inducible, endothelium-specific Cre alleles that can switch conditional null (floxed) target genes off during any required developmental time window, thus circumventing the early embryonic lethality seen with global knockout mice that impairs cardiovascular development (eg, Benedito et al31; Table 2). Likewise, refined inducible overexpression systems can be used to switch genes on at any defined time point during embryonic and postnatal development in the mouse (eg, Wang et al47; Table 2). Combined with novel in vivo models of central nervous system angiogenesis, discussed below, these methods have considerably refined our capacity to decipher molecular mechanisms governing vascular development and to understand the activities of pathways required for vascular development, which are not limited to early embryonic development but extend to organogenesis, maintenance of adult vascular homeostasis, and pathological angiogenesis.Table 2. Mouse Lines Expressing Blood ECs and Lymphatic EC-Specific Cre or a Fluorescent Protein ReporterMouse LineDescription of Mouse LineReferenceEC-specific constitutive Cre-deleter mouse lines Tie1-CreThe Tie1 gene promoter drives Cre expression in these transgenic mice in most embryonic ECs from E8-9 onward but not in all ECs in adult vasculature. There may be expression in some hematopoietic cells.32 Tie2-CreTie2 gene promoter and enhancer sequences linked to Cre drive expression in endothelium through embryogenesis and adulthood. May be expressed also in certain monocyte subpopulation and mesenchymal cells.33 flk1-CreVegfr-2 gene promoter and enhancer sequences driving Cre expression in transgenic mice. Cre is expressed in ECs in embryos from E11.5 or E13.5 onward, depending on the mouse line, except the lung microvasculature.34 VE-cadherin–CreVE-cadherin gene promoter Cre. Early expression in ECs of yolk sac at E7.5; more contiguous EC expression by E14.5 and onward. Shows ubiquitous EC expression in the adult, including LECs and some hematopoietic cells.35LEC-specific constitutive Cre-deleter mouse lines Lyve1-CreGFP-Cre knock in construct in mouse Lyve-1 gene. Cre is expressed in LECs of the lymph nodes but also in a varying fraction of other ECs and in some hematopoietic cells.36EC-specific inducible Cre-deleter mouse lines VE-cadherin–Cre–ERT2VE-cadherin (Cdh5) promoter–Cre–ERT2. Administration of tamoxifen (4-OHT) induces translocation of Cre-ERT2 to the nucleus where it mediates Cre-loxP site-specific combination. Allows EC-specific Cre recombination in a temporally and spatially controlled manner.35, 37 Pdgfb-CreERT2Platelet-derived growth factor B promoter–Cre. Allows EC-specific Cre recombination in most vascular beds of newborn mice but not in all organs in adults (eg, in central nervous system).38LEC-specific Cre-deleter mouse lines Prox1-CreERT2Prox1 gene promoter–driven Cre-ERT2 deletes in the lymphatic vasculature and in other Prox1-expressing cells (eg, in heart and liver), but not in ECs except in venous valves.39, 40Genetic fluorescent protein reporter mouse lines for visualization of lymphatic vessels in living mice Vegfr3-EGFPLucGFP–luciferase fusion protein knock-in construct in mouse Vegfr3 gene, expressed in LECs. Allows fluorescent and luminescent imaging of lymphatic vasculature in embryos and during the first weeks after birth, but expression is strongly downregulated during postnatal development.41 Prox1-GFPFluorescent GFP protein is expressed under the Prox1 promoter in LECs. Recapitulates faithfully the endogenous Prox1 expression. Allows fluorescent imaging of the lymphatic vasculature in embryos and adults.42 Prox1-mOrange2Fluorescent mOrange2 protein expressed under the Prox1 promoter in LECs. Similar expression pattern as in Prox1-GFP mice (Choi et al42). Allows fluorescent imaging of lymphatic vasculature in embryos and adults.43 Prox1-tdTomatoFluorescent tdTomato protein imaging of lymphatic vasculature.44Genetic fluorescent protein reporter mouse lines for visualization of blood vessels in living mice Tie2-GFPTie2 gene promoter and enhancer sequences control GFP expression in transgenic mice. Allows fluorescent imaging of blood vessels during embryogenesis and in adults.45 Tie1-GFPTie1 promoter fragment GFP expression in transgenic mice. GFP is expressed in the ECs in embryos but is downregulated in the adult vasculature.46E indicates embryonic day; EC, endothelial cell; GFP, green fluorescent protein; and LEC, lymphatic endothelial cells.The vertebrate central nervous system is made up of brain, spinal cord, and retina. All 3 organs are vascularized during embryonic and early postnatal development to provide oxygen and nutrients to both neuronal progenitors and the metabolically highly active and terminally differentiated neurons that relay information. Both brain and spinal cord acquire a vascular network before birth, although the complexity of this network increases after birth for some time. In contrast, the retina is initially supplied by the extraretinal choroidal vasculature and hyaloid arteries. Although the choroidal vasculature persists into adulthood, the hyaloid arteries are replaced during postnatal development with an intraretinal vascular system that is known as the retinal vasculature. Brain, spinal cord, and retinal vasculature all form through angiogenesis (reviewed by Ruhrberg and Bautch48). Because of the stereotypic nature of vessel sprouting in all 3 organs, they are extremely well suited to identify even subtle changes caused by the perturbation of environmental or genetic factors that regulate angiogenesis. This section summarizes the advantages and limitations of the mouse embryo hindbrain and postnatal retina as model systems to study physiological angiogenesis and introduces the perinatal retina as an additional model to identify factors that modulate pathological neovascularization. Importantly, all 3 experimental models have been described with much detail in recent publications and are therefore easily applied and suited to standardization of angiogenesis research between different investigators.37,49,502.1.1. Brief Description of Hindbrain VascularizationThe mouse hindbrain is vascularized before birth.48 The vascularization of the mouse embryonic hindbrain begins about E9.5, when vessels sprout into the hindbrain from a perineural vascular plexus. These first hindbrain vessels grow perpendicular to the hindbrain surface toward the ventricular zone (radial vessels) but then change direction to sprout parallel to the ventricular hindbrain surface and anastomose with each other (subventricular vascular plexus; Figure 1A). As the hindbrain tissue thickens as a result of the addition of neurons and the formation of glia, sprouting and fusion move to deeper brain layers.Download figureDownload PowerPointFigure 1. Hindbrain and retina models. Time course of blood vessel ingression into the mouse embryo hindbrain and the perinatal mouse retina. A, Vessels sprout from the perineural vascular plexus (PNVP) into the hindbrain at around embryonic day (E) 9.75 in the mouse and then grow radially toward the ventricular zone. Radial vessels do not invade the subventricular zone but sprout laterally and then anastomose to form a subventricular vascular plexus (SVP) by E12.5. B, Retinal vascularization proceeds from center to periphery in a radial manner during the first week of life and leads to an extensively remodeled superficial plexus composed of capillaries, arteries, and veins. Modified from Ruhrberg and Bautch.482.1.1.1. Advantages of the Hindbrain Model to Study Sprouting AngiogenesisPermits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that compromise viability or result in postmidgestation embryonic lethality; when combined with inducible Cre-LoxP technology, can be extended to the analysis of mouse strains with premidgestation lethality.Robust method for whole-mount labeling of ECs and interacting cell types that can be combined with high-resolution imaging for reliable quantification of angiogenic sprouting, network density, and vessel caliber ECs.Enables the analysis of blood vessel growth within a natural but relatively simple multicellular microenvironment where ECs interact with non-ECs to refine the 3D organ architecture (fewer cell types compared with retinal angiogenesis, ie, neural progenitors, differentiating neurons, and microglia precursors but not glia, complex neural networks, or inflammatory cells).2.1.2. Retinal VascularizationThe mouse retina is vascularized only after birth.38,51 The formation of the retinal vasculature begins on the day of birth, when vessels emerge from the optic nerve head, sprout perpendicularly over the" @default.
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• W1993235364 modified "2023-10-08" @default.
• W1993235364 title "State-of-the-Art Methods for Evaluation of Angiogenesis and Tissue Vascularization" @default.
• W1993235364 cites W143420532 @default.
• W1993235364 cites W154322005 @default.
• W1993235364 cites W1544053957 @default.
• W1993235364 cites W1588633184 @default.
• W1993235364 cites W1599842971 @default.
• W1993235364 cites W1605661308 @default.
• W1993235364 cites W1613258091 @default.
• W1993235364 cites W164687564 @default.
• W1993235364 cites W1679229092 @default.
• W1993235364 cites W1835091344 @default.
• W1993235364 cites W1844495809 @default.
• W1993235364 cites W1919923011 @default.
• W1993235364 cites W1967552160 @default.
• W1993235364 cites W1968078293 @default.
• W1993235364 cites W1968596406 @default.
• W1993235364 cites W1971163936 @default.
• W1993235364 cites W1971175908 @default.
• W1993235364 cites W1973096186 @default.
• W1993235364 cites W1973197740 @default.
• W1993235364 cites W1973938885 @default.
• W1993235364 cites W1976175856 @default.
• W1993235364 cites W1976391205 @default.
• W1993235364 cites W1977059167 @default.
• W1993235364 cites W1978418063 @default.
• W1993235364 cites W1984442770 @default.
• W1993235364 cites W1985907458 @default.
• W1993235364 cites W1986025942 @default.
• W1993235364 cites W1987895314 @default.
• W1993235364 cites W1988126593 @default.
• W1993235364 cites W1988368236 @default.
• W1993235364 cites W1989310884 @default.
• W1993235364 cites W1989440555 @default.
• W1993235364 cites W1990140297 @default.
• W1993235364 cites W1990253071 @default.
• W1993235364 cites W1993681546 @default.
• W1993235364 cites W1993810430 @default.
• W1993235364 cites W1993913999 @default.
• W1993235364 cites W1994028945 @default.
• W1993235364 cites W1995736467 @default.
• W1993235364 cites W1996081149 @default.
• W1993235364 cites W1998877773 @default.
• W1993235364 cites W1999048542 @default.
• W1993235364 cites W1999763539 @default.
• W1993235364 cites W2001236781 @default.
• W1993235364 cites W2003922830 @default.
• W1993235364 cites W2005215351 @default.
• W1993235364 cites W2006912346 @default.
• W1993235364 cites W2006961900 @default.
• W1993235364 cites W2008455196 @default.
• W1993235364 cites W2009516278 @default.
• W1993235364 cites W2010775796 @default.
• W1993235364 cites W2012948722 @default.
• W1993235364 cites W2016400372 @default.
• W1993235364 cites W2017390700 @default.
• W1993235364 cites W2019782175 @default.
• W1993235364 cites W2024343337 @default.
• W1993235364 cites W2024861740 @default.
• W1993235364 cites W2026223672 @default.
• W1993235364 cites W2027654614 @default.
• W1993235364 cites W2029714409 @default.
• W1993235364 cites W2030084515 @default.
• W1993235364 cites W2032156560 @default.
• W1993235364 cites W2033049522 @default.
• W1993235364 cites W2033377265 @default.
• W1993235364 cites W2033558162 @default.
• W1993235364 cites W2034073329 @default.
• W1993235364 cites W2034712955 @default.
• W1993235364 cites W2035106720 @default.
• W1993235364 cites W2035597138 @default.
• W1993235364 cites W2036447543 @default.
• W1993235364 cites W2037921624 @default.
• W1993235364 cites W2038369700 @default.
• W1993235364 cites W2041063291 @default.
• W1993235364 cites W2042130990 @default.
• W1993235364 cites W2042899052 @default.
• W1993235364 cites W2043099950 @default.
• W1993235364 cites W2043379169 @default.

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