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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.
It is now widely accepted that much of the dynamic function of cells and tissues is regulated from outside the cell by the extracellular matrix. In ad- tion to its conventional role in providing a scaffold for building tissues, the extracellular matrix acts as a directional highway for cellular movement and provides instructional information for promoting survival, proliferation, and differentiation. Indeed, the extracellular matrix is beginning to take a starring role in the choreography of cell and tissue function. The diverse roles of the extracellular matrix are reflected in its highly complicated structure, consisting of an ever increasing number of components. Yet the mechanisms of extracellular matrix assembly and how they influences cell behavior are only just beginning to be understood. In order to solve these problems new methodologies are, of necessity, being developed. Many of these technologies are highly sophisticated and are currently available only in a ha- ful of laboratories. However, we believe that they can readily be transported and established by other researchers. Thus, the purpose of Extracellular Matrix Protocols is to present some of these complicated techniques in a style that is relatively easy to reproduce.
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.
Critically acclaimed for more than 25 years, the Methods in Cell Biology series provides an indispensable tool for the researcher. Each volume is carefully edited by experts to contain state-of-the-art reviews and step-by-step protocols. Techniques are described completely so that methods are made accessible to users. This volume, Methods of Cell-Matrix Adhesion, contains integrated coverage on cell-matrix adhesion methods. It brings the classical methodologies and the latest techniques together in one concise volume. This coverage includes experimental protocols and their conceptual background for all aspects of cell-matrix adhesion research: the extracellular matrix, adhesion receptors, and the growing number of functional applications of matrix-adhesion in molecular cell biology. Also covered is the purification of the extracellular matrix to functional analyses of cellular responses.
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.
Methods in Extra Cellular Matrix, Volume 142, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Unique to this updated volume are sections devoted to Elastin, Quantification of collagen and elastin, Fibrillins, Lysyl oxidase, Fibulins, Matrilins, Hyaluronic Acid, Small leucine-rich proteoglycans, Syndecans, Fibronectin, SPARC, Thrombospondins, Tenascins, Collagen IV, Multi-photon analysis of ECM, Cell-derived extracellular matrices, Laminins, Fibrillar Collagens, Imaging ECM in developing embryos, Analysis of Matrix Degradation, Ultrastructural analysis of ECM, Versican and Large proteoglycans, and an ECM crosslink analysis. This series covers a wide array of topics about the extracellular matrix, including an understanding of crucial proteins and glycoproteins components of ECM. Contains contributions from experts in the field from across the world Covers a wide array of topics on the extracellular matrix, including an understanding crucial proteins and the glycoproteins components of ECM Includes analysis based topics, such as quantification of collagen and elastin, mulit-photon analysis of ECM and ECM crosslink analysis
This detailed volume compiles state-of-the-art protocols that will serve as recipes for scientists researching collagen, an abundant protein with great importance to health and disease, as well as in applications like food, cosmetics, pharmaceuticals, cosmetic surgery, artificial skin, and glue. Beginning with a section on in vitro models for the characterization of collagen formation, the book continues by highlighting large-scale analysis of collagen with mass spectrometry in order to elucidate the proteomics, degradomics, interactomes, and cross-linking of collagen, high resolution imaging approaches for collagen by the use of scanning electron microscopy and multiphoton imaging, as well as the role of collagen during physiological and pathological conditions. Written for the highly successful Methods in Molecular Biology 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 practical, Collagen: Methods and Protocols is an ideal guide to high quality and repeatable protocols in this vital field of study.
A versatile collection of readily reproducible cell-cell interaction assays for uncovering cellular interactions at the molecular level, both in vitro and in vivo. The protocols cover a diverse set of cell-cell interaction models in both normal and pathological states, are readily adaptable to nearly any cell type and organ system, and include primary data and outcome analysis. In addition, the protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.