This detailed book builds on the foundation of lymphatic protocols with tools to aid in the interpretation of large datasets created by angiogenesis research. For example, this includes protocols to explore how transcriptional phenotyping can be achieved, both in terms of exploring tissue-specific heterogeneity to the use of database mining of existing datasets. Similarly, bioinformatic techniques run alongside protocols for transcript analysis, cell biology, pathology, as well as physiological and developmental angiogenesis models. Written for the highly successful
series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date,
Angiogenesis: Methods and Protocols provides a meaningful compendium of practical approaches to angio
Table of ContentsPart I: Developmental Biology
1. The Embryonic Mouse Hindbrain Model to Study Sprouting Angiogenesis In Vivo
Gabriela D’Amico and Christiana Ruhrberg
2. A Workflow to Track and Analyze Endothelial Migration during Vascular Development in Zebrafish Embryos Using Lightsheet Microscopy
Yan Chen, Paul C. Evans, and Robert N. Wilkinson
3. Analyzing Vessel Regression and Endothelial Apoptosis as a Component of Angiogenic Vessel Remodeling
Lachlan Whitehead, Emma C. Watson, and Zoe L. Grant
4. Whole-Mount Immunofluorescence Protocol for 3D Imaging, Reconstruction, and Quantification of 4th Pharyngeal Arch Artery Formation
Elena Ioannou and Christiana Ruhrberg
5. Quantifying and Characterizing Angiogenesis Using the Postnatal Mouse Retina
Alessandro Fantin
Part II: Adult Physiology and Pathology
6. Characterization of Lymphatic Vasculature Using Whole-Mount Immunostaining of Mouse Embryonic Dorsal Skin
Nikita Ved and Jacinta Isabelle Kalisch-Smith
7. Analyzing Lymphatic Vessel Patterning in Adult Tissue
Louise A. Johnson
8. Transmission Electron Microscopy of Endothelium
Zubair Ahmed Nizamundeen, Loubna ElSaboni, Grace Vetch, Jumana Karim, Chris R. Neal, Claire L. Allen, David O. Bates, and Kenton P. Arkill
9. Measurement of Revascularisation in the Hind Limb after Experimental Ischemia in Mice
Sohni Ria Bhalla, Federica Riu, Maria J.C. Machado, and David O. Bates
10. Imaging Blood Vessels and Lymphatics in Mouse Trachea Wholemounts
Peter Baluk and Donald M. McDonald
11. Non-Invasive Measurement of Retinal Microvascular Permeability during Loss of Endothelial Quiescence
Naseeb K. Malhi, David O. Bates, Kenton P. Arkill, and Claire L. Allen
12. An Ex Vivo Tissue Culture Method for Discovering Cell Dynamics Involved in Stromal Vascular Fraction Vasculogenesis Using the Mouse Mesentery
Dima Majbour, Ariana D. Suarez-Martinez, Nicholas A. Hodges, Arinola O. Lampejo, Banks M. Lomel, Elijah W. Rice, Hulan Shang, Adam J. Katz, and Walter L. Murfee
13. Quantitative Three-Dimensional Analysis of the Lymphatic Vasculature in the Postnatal Mouse Heart
Konstantinos Klaourakis, Paul R. Riley, and Joaquim Miguel Vieira
14. Quantifying Lymphatic Vessel Density in Human Tissue Samples
Stewart G. Martin, Emad Rakha, and Sarah J. Storr
15. Quantifying Vascular Remodeling in the Mouse Spinal Cord
M.E. Da Vitoria Lobo, Lydia Hardowar, and Richard P. Hulse
16. Quantitative Methods to Assess Adipose Vasculature
Martina Rudnicki, Alexandra Pislaru, and Tara L. Haas
17. Quantification of Angiogenesis in Laser Choroidal Neovascularization
Kathryn R. Green, Nicholas Beazley-Long, Amy P. Lynch, Claire L. Allen, David O. Bates, and Andrew V. Benest
Part III: Reducing Complexity: Growing and Studying Endothelial Cells Outside the Body
18. A Rapid Adaptable Method for Isolation of Endothelial Cells from Human Adipose Tissue
Jacqueline Herold and Joanna Kalucka
19. Simple Gene Knockdown in Endothelial Cells Using Short Interfering RNA Oligonucleotides
Zarah B. Tabrizi, Kathryn R. Green, Amy P. Lynch, Nada S. Ahmed, Nicholas Beazley-Long, and Andrew V. Benest
20. Transcription Factor Chromatin Immunoprecipitation in Endothelial Cells
Philip Kitchen, Kevin Gaston, and Padma-Sheela Jayaraman
21. In Vitro Co-Culture of Fibroblast and Endothelial Cells to Assess Angiogenesis
Adam F. Odell and Aarren J. Mannion
22. Quantification of Leukocyte-Endothelial Cell Interaction during In Vitro Models of Thrombosis at Arterial and Venous Shear Rates
Joshua H. Bourne
23. Isolation of Cardiac Endothelial Cells for Transcriptomic Analysis of the Zebrafish and Mouse Heart
Ziwen Li, Katherine M. Ross Stewart, Finnius A. Bruton, Martin A. Denvir, and Mairi Brittan
24. The Evaluation of Neovessel Angiogenesis Behavior at Tissue Interfaces
Hannah A. Strobel and James B. Hoying
25. Endothelial Cell Fibrin Gel Angiogenesis Bead Assay
Eva Clavane, Hannah Taylor, Richard Cubbon, and Paul Meakin
26. Live Cell Imaging and Analysis of Cancer-Cell Transmigration through Endothelial Monolayers
Aarren J. Mannion
27. Differentiation and Characterization of Human Pluripotent Stem Cell-Derived Cardiac Endothelial Cells for In Vitro Applications
Kavita Raniga, Nguyen T.N. Vo, and Chris Denning
Part IV: Making Use of Publically Existing Datasets and Resources
28. Finding and Verifying Enhancers for Endothelial-Expressed Genes
Alice Neal, Helena Rodriguez-Caro, and Sarah De Val
29. Endothelial Cell RNA-Seq Data: Differential Expression and Functional Enrichment Analyses to Study Phenotypic Switching
Guillermo Díez Pinel, Joseph L. Horder, John R. King, Alan McIntyre, Nigel P. Mongan, Gonzalo Gómez López, and Andrew V. Benest
