Angiogenesis Assays: A Critical Appraisal of Current Techniques describes in detail the in vitro and in vivo assays currently being used to study angiogenesis, including the provision of protocols.
The chapters are written by leading scientists who use these techniques in their angiogenesis research programs and who deliver here a critical appraisal of their strengths and weaknesses.
Divided into three sections:
- in vitro assays of endothelial cell activity
- various in vivo angiogenesis assays
- some of the biological endpoints being used in clinical trials to assess the clinical efficacy of anti-angiogenic drugs
Contents
Endothelial cell biology
- Morphology of the endothelium
- Endothelial cell adhesion and interactions
- Coagulation and haemostasis
- Transport
- Angiogenesis
- Isolation and culture of endothelial cells
- Endothelial cell heterogeneity and organ specificity
- Gene expression in endothelial cells
Endothelial cell proliferation assays
- Cell proliferation
- Cell proliferation assays
Endothelial cell migration assays
- Transfilter assay
- 'Wound healing' assay
- Teflon fence assay
- Phagokinetic track assay
- In/ex vivo approaches for studying endothelial cell migration
Tubule formation assays
- Endothelial cell sources
- Endothelial cell morphology and tubule formation
- Tubule assay matrices
- 2D assay protocols
- Analysis of tubule formation in 2D assays
- Recent developments in 2D assays
- 3D assays
- Analysis of 3D assays
- Co-culture assays
Modelling the effects of the haemodynamic environment on endothelial cell responses relevant to angiogenesis
- Definitions
- Experimental patterns of shear exposure and relevance to 'angiogenesis' assays
- Methods for studying responses of endothelial cells exposed to shear stress
- Readouts in flow cultures relevant to angiogenesis
- Critical considerations and conclusions
Whole or partial vessel outgrowth assays
- Rat aortic ring assay
- Mouse aorta models
- Chick aortic arch assay
- Porcine carotid artery assay
- Human explant cultures
- Recent developments
- Quantification
- Strengths and weaknesses of organ culture assays
Assaying endothelial–mural cell interactions
- 2D models to study endothelial–mural cell interactions
- 3D models to study blood vessel assembly
Assays for membrane and intracellular signalling events
- The endothelial caveolae as the organizers of efficient spatial signal transduction mechanisms
- The prostaglandins forming system: regulation of signal transduction and angiogenesis
- Use of antisense oligo and sirna to evaluate the function of single signalling molecules
- Knocking down and knocking out Caveolin-1 gene
- Analysing protein-protein interactions in signalling molecules involved in angiogenesis
Implantation of sponges and polymers
- Materials used in implantation techniques
- Implantation technique for assessment of inflammatory processes
- Implantation technique for assessment of systemic pathological conditions
- Implantation technique for assessment of tumour angiogenesis
- Implantation technique for assessment of inflammatory angiogenesis in genetically modified mice
- Applications of cannulated sponge model for testing angiogenesis modulators
- Techniques for assessment of the vascularization in implants
- Summary of cannulated sponge assay: advantages and disadvantages
Angiogenesis assays in the chick
- Overview of the basic 'in shell' CAM assay
- Variations on the CAM assay
- Summary of the CAM assay: disadvantages and advantages
Corneal angiogenesis assay
- Anatomy and histology
- Brief history of CNV assays
- The process of corneal neovascularization
- Experimental induction of corneal neovascularization
Dorsal air sac model
- 12.2 Preparation of the DAS model
- evaluation of anti-angiogenic photodynamic therapy using a DAS model
- Determination of blood volume or blood flow in the angiogenic site using the DAS model
- Quantitative or semi-quantitative analysis of angiogenesis in the DAS model
- Isolation and application of neovessel-targeting probe using DAS model
Chamber assays
- In vivo imaging of angiogenesis and microcirculation
- Chamber assays
- Physiological angiogenesis
- Tumour angiogenesis
- Angiogenesis in endometriosis
- Angiogenesis in wound and bone healing
- Angiogenesis in ischaemia and hypoxia
- Angiogenesis in transplantation
- Angiogenesis in biomaterial incorporation
- Angiogenesis in tissue engineering
- Conclusions and perspectives
Tumour models: analysis of angiogenesis in vivo
- Tumour microenvironment and angiogenesis
- Tumour models
- Subcutaneous tumour models
- Orthotopic tumour models
- Transgenic mouse models of tumour angiogenesis
- Monitoring vascular permeability
- Analysis of angiogenesis in tumours
Angiomouse: imageable models of angiogenesis
- Fluorescent proteins to image angiogenesis
- Dual-colour tumour host models
- ND-GFP mouse model
- Methods of angiogenesis analysis in GFP models
Techniques and advances in vascular imaging in Danio rerio
- Visualizing the developing vasculature
- In situ hybridization reveals vascular specific expression
- Vascular specific fluorescent reporter lines
- Chemical mutagenesis screens reveal novel genes involved in cardiovascular development
Biological and clinical implications of recruitment of stem cells into angiogenesis
- Phenotypic and functional characterization of EPCS
- Mobilization, homing and differentiation of EPCS
- Ageing
- Therapeutic applications
Methods for monitoring of the anti-angiogenic activity of agents in patients: novel trial design
- Tumour micro-environmental factors
- Possible mechanisms of acquired resistance to anti-angiogenic drugs
- Standard chemotherapy versus angiogenesis inhibitors
- Monitoring clinical trials
- Tumour dormancy and tumour progression
- Tumour markers
- Microvessel density
- Surrogate endpoints
- Molecular imaging
- New insights into trial design
An overview of current angiogenesis assays: Choice of assay, precautions in interpretation, future requirements and directions
- In vitro assays
- In vivo assays
- Clinical trials
Index
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