Main description:

This volume details established experimental protocols on the isolation, characterization, and detection of extracellular matrix (ECM) molecules as well as methods to study the activity and role of ECM components on various biological functions, the formation of exosomes, and tissue surrogates. Chapters display a variety of protocols ranging from biochemical, cell, and molecular biology assays to complex tissue imaging techniques and in vivo models to elucidate the role of the extracellular matrix. Written in the highly successful Methods in Molecular Biology series format, 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 cutting-edge, The Extracellular Matrix: Methods and Protocols aims to provide protocols and applications that can be used by both Ph.D. students and senior researchers to advance research in the field of extracellular matrix biology.

Contents:

1. The Complex Interplay between Extracellular Matrix and Cells in Tissues

Dimitra Manou, Ilaria Caon, Panagiotis Bouris, Irene-Eva Triantaphyllidou, Cristina Giaroni, Alberto Passi, Nikos K. Karamanos, Davide Vigetti, and Achilleas D. Theocharis

2. Method for Studying ECM Expression: In Situ RT-PCR

Elena Carava, Cristiana Marcozzi, Barbara Bartolini, Marcella Reguzzoni, Paola Moretto, Ilaria Caon, Evgenia Karousou, Alberto Passi, and Manuela Viola

3. Visualizing the Supramolecular Assembly of Collagen

Mario Raspanti, Marcella Reguzzoni, Petra Rita Basso, Marina Protasoni, and Desiree Martini

4. Steady-state and Pulse-chase Analyses of Fibrillar Collagen

Antonella Forlino, Francesca Tonelli, and Roberta Besio

5. Methods for Isolation and Characterization of Sulfated Glycosaminoglycans from Marine Invertebrates

Mariana P. Stelling, Ananda de A. Bento, Philippe Caloba, Eduardo Vilanova, and Mauro S. G. Pavao

6. Analysis of Proteoglycan Synthesis and Secretion in Cell Culture Systems

Chiara Paganini, Rossella Costantini, and Antonio Rossi

7. Analysis of UDP-Sugars from Cultured Cells and Small Tissue Samples

Sanna Oikari and Markku I Tammi

8. Methods for Hyaluronan Molecular Mass Determination by Agarose Gel Electrophoresis

Mary K. Cowman

9. Hyaluronan Pericellular Matrix: Particle Exclusion Assay

Melanie A. Simpson

10. Determination of Cell Surface Hyaluronan through Flow Cytometry

Daiana L. Vitale, Fiorella M. Spinelli, and Laura Alaniz

11. Method for Detecting Hyaluronan in Isolated Myenteric Plexus Ganglia of Adult Rat Small Intestine

Michela Bistoletti, Paola Moretto, and Cristina Giaroni

12. Analyzing Hyaluronidases in Biological Fluids

Christos Velesiotis, Stella Vasileiou , and Demitrios H. Vynios

13. Method for Studying the Physical Effect of Extracellular Matrix on Voltage-dependent Ion Channels Gating

Eleonora Solari and Andrea Moriondo

14. Methods for Monitoring Matrix-Induced Autophagy

Carolyn Chen, Aastha Kapoor, and Renato V. Iozzo

15. Method for Determining Gelatinolytic Activity in Tissue: In Situ Gelatin Zymography

Elin Hadler-Olsen and Jan-Olof Winberg

16. Method for Determining Gelatinolytic Activity in Tissue Extracts: Real-Time Gelatin Zymography

Elin Hadler-Olsen and Jan-Olof Winberg

17. In Vitro Spheroid Sprouting Assay of Angiogenesis

Fatema Tuz Zahra, Efrossini Choleva, Md Sanaullah Sajib, Evangelia Papadimitriou, and Constantinos . Mikelis

18. Matrigel Plug Assay for In Vivo Evaluation of Angiogenesis

Pinelopi Kastana, Fatema Tuz Zahra, Despoina Ntenekou, Stamatiki Katraki-Pavlou, Dimitris Beis, Michail S. Lionakis, Constantinos . Mikelis, and Evangelia Papadimitriou

19. Exosomes from Cell Culture Conditioned Medium: Isolation by Ultracentrifugation and Characterization

Anurag Purushothaman

20. Preparation and Characterisation of Rich in Extracellular Matrix Tissue Surrogates Using the Principles of Macromolecular Crowding

Adrian Djalali-Cuevas, Sergio Garnica-Galvez, Andrea Rampin, Diana Gaspar, Ioannis Skoufos, Athina Tzora, Nikitas Prassinos, Nikolaos Diakakis, and Dimitrios I. Zeugolis

21. Novel Approaches for Extracellular Matrix Targeting in Disease Treatment

Nikolaos A. Afratis and Irit Sagi