Adams Methods in Cell-Matrix Adhesion

1. Auflage 2002
ISBN: 978-0-08-052496-2
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band Volume 69, 461 Seiten, Web PDF

Reihe: Methods in Cell 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.

Autoren/Hrsg.

Adams, Josephine

Inhaltsverzeichnis

1;Cover;1
2;Copyright Page;5
3;Contents;6
4;Contributors;12
5;Acknowledgments;16
6;Part I: Preface and Perspectives on Cell–Matrix Adhesion;18
6.1;Preface;20
6.2;Cell–Matrix Interactions: The View from the Outside;24
6.3;Matrix and Meaning;30
6.4;Cell–Matrix Adhesion Research and the Development of Biotherapeutics;34
7;Part II: Matrix Methodologies;42
7.1;Chapter 1. Detection and Purification of Instructive Extracellular Matrix Components with Monoclonal Antibody Technologies;44
7.1.1;I. Introduction;45
7.1.2;II. Methods for Detecting ECM Components That Instruct Cell Responses in Reporter Cells;49
7.1.3;III. Preparation of Mabs for Identification of Instructive ECM;58
7.1.4;IV. Purification of Instructive ECM with Mabs and Interactions with Reporter Cells;64
7.1.5;V. Summary;66
7.1.6;References;66
7.2;Chapter 2. Isolation and Purification of Proteoglycans;70
7.2.1;I. Overview and Scope;70
7.2.2;II. Extraction of Proteoglycans;72
7.2.3;III. Radiolabeling of Proteoglycans for Ease of Purification;74
7.2.4;IV. Anion-Exchange Chromatography;74
7.2.5;V. Hydrophobic Interaction Chromatography;76
7.2.6;VI. Gel Filtration Chromatography;77
7.2.7;VII. Immunoaffinity Chromatography;78
7.2.8;VIII. Quantification and Characterization;80
7.2.9;References;83
7.3;Chapter 3. Expression of Recombinant Matrix Components Using Baculoviruses;86
7.3.1;I. Introduction: The Importance of Recombinant Extracellular Matrix Molecules for Protein Studies;87
7.3.2;II. Requirements and Rationale for Development of a General Protein Expression System;87
7.3.3;III. Expression in Insect Cells Using pCOCO;91
7.3.4;IV. Results Obtained from the COCO System;95
7.3.5;References;97
7.4;Chapter 4. Heparan Sulfate–Growth Factor Interactions;100
7.4.1;I. Introduction: HS Regulation of Growth Factor Signaling;100
7.4.2;II. Methods for Identifying the Role of HS in Growth Factor Binding and Signaling;106
7.4.3;III. Identification of Regulatory HS;112
7.4.4;IV. Analyzing the Structure of HS: General Considerations and Methodologies;119
7.4.5;References;121
7.5;Chapter 5. Analysis of Basement Membrane Self-Assembly and Cellular Interactions with Native and Recombinant Glycoproteins;128
7.5.1;I. Introduction;128
7.5.2;II. Native and Recombinant Basement Membrane Proteins;131
7.5.3;III. Analysis of Self-Assembly in Vitro;141
7.5.4;IV. Analysis of Cell-Surface Contributions to Basement Membrane Assembly;152
7.5.5;V. Approaches to Evaluating ECM Molecules for Assembly Interactions;153
7.5.6;VI. Integration of Genetic and Molecular Approaches;154
7.5.7;References;155
7.6;Chapter 6. Preparation and Analysis of Synthetic Multicomponent Extracellular Matrix;162
7.6.1;I. Introduction to the ECM;162
7.6.2;II. Development of Synthetic Matrices;164
7.6.3;III. Discussion;172
7.6.4;References;175
7.7;Chapter 7. Analysis of Matrix Dynamics by Atomic Force Microscopy;180
7.7.1;I. Introduction;181
7.7.2;II. General Experimental Design;182
7.7.3;III. Probe Microscopy in Fluid: AFM Imaging;185
7.7.4;IV. Probe Microscopy in Fluid: Molecular Pulling/Force Spectroscopy;191
7.7.5;V. Probe Microscopy in Air: AFM Imaging;199
7.7.6;VI. Conclusion;205
7.7.7;References;205
7.8;Chapter 8. Analysis of Matrix Degradation;212
7.8.1;I. Introduction;212
7.8.2;II. Experimental Approaches and Protocols;213
7.8.3;References;221
8;Part III: Adhesion Receptor Methodologies;224
8.1;Chapter 9. Expression Cloning Strategies for the Identification of Adhesion Molecules;226
8.1.1;I. Introduction;226
8.1.2;II. Importance of Methodology;229
8.1.3;III. General Considerations;229
8.1.4;IV. Protocols;231
8.1.5;V. Points of Variation;235
8.1.6;VI. Critical Controls;236
8.1.7;VII. Potential Use in Disease/Drug Screening Applications;236
8.1.8;References;237
8.2;Chapter 10. Purification of Integrins and Characterization of Integrin-Associated Proteins;240
8.2.1;I. Introduction;241
8.2.2;II. Isolation of Integrins: General Considerations and Points of Variation;242
8.2.3;III. Characterization of Integrin-Associated Proteins through mAb Production: General Considerations and Points of Variation;251
8.2.4;References;258
8.3;Chapter 11. Methods for Analysis of the Integrin Ligand Binding Event;264
8.3.1;I. Introduction;264
8.3.2;II. Application of Affinity Cross-Linking to Study of the Integrin Ligand Binding Pocket;265
8.3.3;III. Microtiter-Based Integrin Binding Assays;267
8.3.4;IV. Measuring Ligand Binding to Integrins with Surface Plasmon Resonance;271
8.3.5;V. Analyzing Integrin Redox Status;273
8.3.6;References;275
8.4;Chapter 12. Intracellular Coupling of Adhesion Receptors: Molecular Proximity Measurements;278
8.4.1;I. Introduction and Applications;278
8.4.2;II. Fluorescent Lifetime Imaging Microscopy (FLIM);282
8.4.3;References;293
9;Part IV: Functional Applications of Cell–Matrix Adhesion in Molecular Cell Biology;296
9.1;Chapter 13. Functional Analysis of Cell Adhesion: Quantitation of Cell–Matrix Attachment;298
9.1.1;I. Introduction;298
9.1.2;II. General Concepts in Cell Adhesion Assays;300
9.1.3;III. Cell Attachment Assays;303
9.1.4;IV. Cell Spreading Assays;305
9.1.5;V. Direct Binding Assays;306
9.1.6;VI. Cell Migration Assays;307
9.1.7;VII. Conclusions;310
9.1.8;References;310
9.2;Chapter 14. Measurements of Glycosaminoglycan-Based Cell Interactions;314
9.2.1;I. Introduction;314
9.2.2;II. Cell Adhesion Assay;316
9.2.3;III. Cell Invasion Assay;319
9.2.4;References;324
9.3;Chapter 15. Applications of Adhesion Molecule Gene Knockout Cell Lines;326
9.3.1;I. Introduction;327
9.3.2;II. Derivation of Somatic Cell Lines;328
9.3.3;III. ES Cell Lines Carrying Homozygous Mutations of Integrin Adhesion Molecule Genes;332
9.3.4;IV. Studies with Adhesion Receptor Null Cell Lines;334
9.3.5;V. Studies with Extracellular Matrix Component Null Cell Lines;338
9.3.6;VI. Summary;339
9.3.7;References;339
9.4;Chapter 16. Flexible Polyacrylamide Substrata for the Analysis of Mechanical Interactions at Cell–Substratum Adhesions;342
9.4.1;I. Introduction;343
9.4.2;II. Preparation of the Polyacrylamide Substratum;344
9.4.3;III. Characterization of the Gel;347
9.4.4;IV. Data Collection for the Analysis of Traction Forces;349
9.4.5;V. Analysis of Traction Forces;351
9.4.6;VI. Other Applications Using Polyacrylamide Substrata;353
9.4.7;VII. Summary;355
9.4.8;References;355
9.5;Chapter 17. Cell Migration in Slice Cultures;358
9.5.1;I. Introduction;358
9.5.2;II. Somitic Migration;359
9.5.3;III. Migration from the Subventricular Zone to the Olfactory Bulb;366
9.5.4;IV. Axonal Outgrowth;370
9.5.5;V. Conclusions and Perspectives;373
9.5.6;References;374
9.6;Chapter 18. Application of Cell Adhesion to Study Signaling Networks;376
9.6.1;I. Introduction;376
9.6.2;II. Applications;378
9.6.3;III. Methods;379
9.6.4;References;399
9.7;Chapter 19. Use of Micropatterned Adhesive Surfaces for Control of Cell Behavior;402
9.7.1;I. Introduction;402
9.7.2;II. Materials and Instrumentation;409
9.7.3;III. Procedures;411
9.7.4;IV. Comments;416
9.7.5;V. Pitfalls;416
9.7.6;References;417
9.8;Chapter 20. Adenoviral-Mediated Gene Transfer in Two-Dimensional and Three-Dimensional Cultures of Mammary Epithelial Cells;420
9.8.1;I. Introduction;421
9.8.2;II. Using Collagen I as a Three-Dimensional Matrix for Cell Culture;422
9.8.3;III. Using Reconstituted Basement Membrane as a Three-Dimensional Matrix for Cell Culture;424
9.8.4;IV. Preparing Mouse Mammary Epithelial Cell Cultures;426
9.8.5;V. Preparing Recombinant Adenovirus for Infecting Mammary Epithelial Cells;426
9.8.6;VI. Infecting Mammary Epithelial Cell Cultures with Adenovirus;436
9.8.7;References;438
10;Part V: General Information;442
10.1;Appendix A: List of Suppliers;444
10.2;Appendix B: Relevant Microarray Dataset Experiments;448
10.3;Appendix C: Web Site Resources of Interest;452
11;Index;458
12;Volumes in Series;472