Fibrosis Research Methods and Protocols

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™ 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.

The Clinical Scope of Fibrosis.- Pulmonary Fibrosis.- Renal Fibrosis.- Transforming Growth Factor-?.- Regulation of Extracellular Matrix Metabolism: Cultured Cells.- Isolation and Culture of Skin Fibroblasts.- Isolation and Culture of Hepatic Stellate Cells.- Isolation and Phenotypic Characterization of Lung Fibroblasts.- Methods for Measuring Type I Collagen Synthesis In Vitro.- Methods for Assessing the Molecular Mechanisms Controlling Gene Regulation.- Methods for Measuring TGF-? Using Antibodies, Cells, and Mice.- Studying Fibrosis Using Animal Models.- Morphological Methods for Assessment of Fibrosis.- Methods for Measuring Hydroxyproline and Estimating In Vivo Rates of Collagen Synthesis and Degradation.- Approaches to Evaluation of Fibrogenic Pathways in Surgical Lung Biopsy Specimens.- Animal Models for Adult Dermal Wound Healing.- Modeling Liver Fibrosis in Rodents.- Animal Models of Pulmonary Fibrosis.- Animal Models of Renal Fibrosis.- Animal Models of Cardiac Fibrosis.- Genetic Approaches to Studying Fibrosis.- Finding Fibrosis Genes.- Genetic Studies to Identify Hepatic Fibrosis Genes and SNPs in Human Populations.- Analysis of Microarray Experiments for Pulmonary Fibrosis.- DNA Microarrays and Data Mining To Study Hepatic Fibrosis.- Analysis of Proteins Dominantly Expressed in Hepatic Stellate Cells of Activated Phenotype.