Part I. The Clinical Scope of Fibrosisn n Pulmonary Fibrosisn David A. Zisman, Michael P. Keane, John A. Belperio, Robert M. Strieter, and Joseph P. Lynch, IIIn n Renal Fibrosisn H. William Schnapern n Transforming Growth Factor-b: A Key Mediator of Fibrosisn Alain Mauvieln n Part II. Regulation of Extracellular Matrix Metabolism: Cultured Cellsn n Isolation and Culture of Skin Fibroblastsn Laure Rittié and Gary J. Fishern n Isolation and Culture of Hepatic Stellate Cellsn Ralf Weiskirchen and Axel M. Gressnern n Isolation and Phenotypic Characterization of Lung Fibroblastsn Carolyn J. Baglole, Sireesha Y. Reddy, Stephen J. Pollock, Steven E. Feldon, Patricia J. Sime, Terry J. Smith, and Richard P. Phippsn n Methods for Measuring Type I Collagen Synthesis In Vitron David C. Rishikof, Ping-Ping Kuang, Mangalalaxmy Subramanian, and Ronald H. Goldsteinn n Methods for Assessing the Molecular Mechanisms Controlling Gene Regulationn Richard A. Rippe and Branko Stefanovicn n Methods for Measuring TGF-b Using Antibodies, Cells, and Micen Vladimir Jurukovski, Branka Dabovic, Vesna Todorovic, Yan Chen, and Daniel B. Rifkinn n Part III. Studying Fibrosis Using Animal Modelsn n Morphological Methods for Assessment of Fibrosisn Rakesh K. Kumarn n Methods for Measuring Hydroxyproline and Estimating In Vivo Rates of Collagen Synthesis and Degradationn Robin J. McAnultyn n Approaches to Evaluation of Fibrogenic Pathways in Surgical Lung Biopsy Specimensn Cory M. Hogaboam, Kristin J. Carpenter, Holly Evanoff, and Steven L. Kunkeln n Animal Models for Adult Dermal Wound Healingn Mary Birch, Annette Tomlinson, and Mark W. J. Fergusonn n Modeling Liver Fibrosis in Rodentsn Christothea Constandinou, Neil Henderson, and John P. Iredalen n Animal Models of Pulmonary Fibrosisn MehrnazGharaee-Kermani, Matthew Ullenbruch, and Sem H. Phann n Animal Models of Renal Fibrosisn Michael Zeisberg, Mary A. Soubasakos, and Raghu Kallurin n Animal Models of Cardiac Fibrosisn Yao Sun and Karl T. Webern n Part IV. Genetic Approaches to Studying Fibrosisn n Finding Fibrosis Genes: The Lungn Lauranell H. Burch and David A. Schwartzn n Genetic Studies to Identify Hepatic Fibrosis Genes and SNPs in Human Populationsn Christopher Paul Dayn n Analysis of Microarray Experiments for Pulmonary Fibrosisn Nilesh B. Davé and Naftali Kaminskin n DNA Microarrays and Data Mining to Study Hepatic Fibrosisn Bernd Schnabl, Youkyung Hwang Choi, Curt H. Hagedorn, and Ramón Batallern n Analysis of Proteins Dominantly Expressed in Hepatic Stellate Cells of Activated Phenotype: The Molecular Approach for Liver Fibrosisn Norifumi Kawadan n Index
Leading investigators review the highlights of current fibrosis research and the experimental methodologies used uncover the mechanisms that drive it. In their discussion of research methodologies utilizing cultured cells to model various aspects of the fibrotic response in vitro, the authors describe the isolation, characterization, and propagation of mesenchymal cells, and highlight the similarities and differences between methods that are appropriate for different types of fibroblasts. Approaches for studying collagen gene regulation and TGF-b production are also discussed, along with experimental methodologies utilizing animal models to study the pathogenesis of fibrosis. The protocols follow the successful Methods in Molecular Medicine(TM) 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.