Extracellular Matrix Protocols

Expert researchers explore this developing field and provide techniques which can be reproduced in most cell biology labs. This new edition contains cutting-edge methods that are worth publicizing in the field and may encourage collaboration with expert labs.

The study of the extracellular matrix (ECM) and its diverse roles in tissue scaffolding and cellular signaling in both physiological and pathological processes has significantly expanded over the past decade. Although well appreciated, the structural and biochemical complexity and the dynamic nature of the living matrix are still under extensive inv- tigation, yielding a growing number of methods with varying degree of sophistication and intricacy. In this edition of Extracellular Matrix Protocols, we compiled a variety of methods that can be readily reproduced in most cell biology laboratories, as well as several cutting-edge technologies that are indeed available in a limited number of centers, but are well worth the awareness and exposure to the ECM research community. As in the previous edition, the book chapters are divided into sections that rep- sent molecular biology techniques to study gene expression, biophysical and biochemical methods for the analysis of structure and composition, cell biology methods for the asse- ment of cell behavior and matrix assembly and tissue engineering applications. All chapters were contributed by scientists who developed the methods or mastered and perfected methods that were routinely used in their laboratories. An effort was made to provide practical working details and helpful notes for the nonexpert user in order to assist reproducibility and accuracy. We hope that these valuable protocols will become helpful tools for ECM researchers and will be further developed and tailored to the s- cific needs of a growing number of applications.

A comprehensive protocol book featuring cutting-edge methods and techniques to study molecular, cellular, biochemical, and biophysical aspects of the extracellular matrix Bench-tested in-vivo, ex-vivo and in-vitro methods written by experts who provided detailed step-by-step instructions and trouble-shooting Easy-to-follow techniques that ensure reproducibility and save time and resources Valuable protocols that can be easily adapted and further modified to fit the individual needs of research teams Includes supplementary material: sn.pub/extras

Klappentext

The study of the extracellular matrix has significantly expanded over the past decade. Although thoroughly researched, the complexity and the dynamic nature of the matrix are still under extensive investigation, yielding a growing number of methods of analysis. In Extracellular Matrix Protocols: Second Edition, expert researchers explore this developing field and provide a number of techniques which can be reproduced in most cell biology labs. This edition contains methods that represent cutting edge unique technologies which are well worth exposure to the broad field and may encourage collaboration with expert labs. Chapters include information on molecular biology techniques to study gene expression, biophysical and biochemical methods for the analysis of structure and composition, and cell biology methods for assessment of cell behavior, matrix assembly and tissue engineering applications. Composed in the highly successful Methods in Molecular Biology(tm) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Extracellular Matrix Protocols: Second Edition investigates the diverse roles of the extracellular matrix, providing important tools for all researchers in the ECM community.

Inhalt
Molecular Biology.- Retroviral delivery of ECM genes.- Tissue-Specific KO of ECM Proteins.- Recombinant Collagen Trimers from Insect Cells and Yeast.- Eukaryotic Expression and Purification of Recombinant Extracellular Matrix Proteins Carrying the Strep II Tag.- Preparation of recombinant fibronectin fragments for functional and structural studies.- Quantitative Determination of Collagen Cross-links.- Biochemical and biophysical analysis.- ECM Macromolecules: Height-mapping and Nano-mechanics Using Atomic Force Microscopy.- Atomic Force Microscopy Measurements of Intermolecular Binding Forces.- Mass-Mapping of ECM Macromolecules by Scanning Transmission Electron Microscopy.- Chemical Microscopy of Biological Samples by Dynamic Mode Secondary Ion Mass Spectrometry (SIMS).- ECM Macromolecules: Rotary Shadowing and Transmission Electron Microscopy.- Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Substrates.- Solid Phase Assays for Studying ECM ProteinProtein Interactions.- Cell Adhesion Assays.- Cell biology assays.- ECM Degradation Assays for Analyzing Local Cell Invasion.- Fluorescence-Based Assays for In Vitro Analysis of Cell Adhesion and Migration.- Fibrin Gel Model for Assessment of Cellular Contractility.- Fluorescent Labeling Techniques for Investigation of Fibronectin Fibrillogenesis (Labeling Fibronectin Fibrillogenesis).- Stromagenesis During Tumorigenesis: Characterization of Tumor-associated Fibroblasts and Stroma-derived 3D Matrices.- Tissue Recombinants to Study Extracellular Matrix Targeting to Basement Membranes.- Organ models.- ECM and FGF-Dependent Assay of Embryonic SMG Epithelial Morphogenesis: Investigating Growth Factor/Matrix Regulation of Gene Expression During Submandibular Gland Development.- Analyzing how Cell AdhesionControls Mammary Gland Function by Transplantation of Embryonic Mammary Tissue from Knockout Mice.- Tissue Engineering.- Characterizing ECM Production by Cells Encapsulated in Hydrogels.- Tissue Engineering and Cell-Populated Collagen Matrices.