1. Human Fibrotic Diseases: Current Challenges in Fibrosis Research Joel Rosenbloom, Edward Macarak, Sonsoles Piera-Velazquez, and Sergio A. Jimenez
Part II: Animal Models of Fibrosis 2. The Bleomycin Model of Pulmonary Fibrosis Tianju Liu, Francina Gonzalez De Los Santos, and Sem H. Phan
3. Intra-Dermal Injections of Bleomycin to Model Skin Fibrosis Toshiyuki Yamamoto
4. Assessing the Effects of Fibrosis on Lung Function by Light Microscopy-Coupled Stereology Bartosz Pilecki and Grith Lykke Sorensen
5. Transplanting Human Skin Grafts onto Nude Mice to Model Skin Scars Jie Ding and Edward E. Tredget
6. Hypertrophic Scarring in the Rabbit Ear: A Practical Model for Studying Dermal Fibrosis Layla Nabai and Aziz Ghahary
7. Mouse and Rat Models of Induction of Hepatic Fibrosis and Assessment of Portal Hypertension Sabine Klein, Robert Schierwagen, Frank Erhard Uschner, and Jonel Trebicka
8. Mouse Models of Corneal Scarring Laure Rittié, Audrey E.K. Hutcheon, and James D. Zieske
9. Modeling Cardiac Fibrosis in Mice: (Myo)Fibroblast Phenotype after Ischemia Daniela Fraccarollo, Paolo Galuppo, and Johann Bauersachs
10. Characterization of Mesenchymal-Fibroblast Cells using the Col1a2 Promoter/Enhancer Ian M. H. Li, Amy L. Horwell, Grace Chu, Benoit de Crombrugghe, and Goerge Bou-Gharious<
Part III: Cell Culture Models
11. Isolation and Culture of Primary Murine Hepatic Stellate Cells Sabine Weiskirchen, Carmen G. Tag, Sibille Sauer-Lehnen, Frank Tacke, and Ralf Weiskirchen
12. Isolation and Culture of Adipose-Derived Stromal Cells from Sub-Cutaneous Fat Lawrence J. Liew, Huan Ting Ong, and Rodney J. Dilley
13. Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting Tripp Leavitt, Michael S. Hu, and Michael T. Longaker
14. Detection of Infiltrating Mast Cells using a Modified Toluidine Blue Staining Nahum Puebla-Osorio, Seri N. E. Sarchio, Stephen E. Ullrich, and Scott N. Byrne
16. Traction Force Measurement using Deformable Microposts Tianfa Xie, Jamar Hawkins, and Yubing Sun
17. Mechanical Deformation of Cultured Cells with Hydrogels Christal A. Worthen, Laure Rittié, and Gary J. Fisher
18. Preparation of Decellularized Lung Matrices for Cell Culture and Protein Analysis Franziska E. Uhl, Darcy E. Wagner, and Daniel J. Weiss
Part IV: ECM Proteins -- Purification, Quantification, and Analysis
19. Type I Collagen Purification from Rat Tail Tendons Laure Rittié
20. Purification of Human Plasma/Cellular Fibronectin and Fibronectin Fragments Giampiero Pietrocola, Simonetta Rindi, Giulia Nobile, and Pietro Speziale
21. Laser Capture Microdissection of Tissue Sections for High-Throughput RNA Analysis Julia Hoffmann, Jochen Wilhelm, and Grazyna Kwapiszewska
22. Collagen Quantification in Tissue Specimens João Quintas Coentro, Héctor Capella-Monsonís, Valeria Graceffa, Zhuning Wu, Anne Maria Mullen, Michael Raghunath, and Dimitrios I. Zeugolis
23. Methods for the Assessment of Active Transforming Growth Factor-β in Cells and Tissues Alison E. John, Joanne Porte, Gisli Jenkins, and Amanda L. Tatler
24. Visualizing in Vitro Type I Collagen Fibrillogenesis by Transmission Electron Microscopy J. Robin Harris
25. Histological and Electron Microscope Staining for the Identification of Elastic Fiber Networks Elaine C. Davis and Ling Li
26. Method for Picrosirius Red-Polarization Detection of Collagen Fibers in Tissue Sections Laure Rittié
27. Probing Collagen Organization: Practical Guide for Second Harmonic Generation (SHG) Imaging Riccardo Cicchi and Francesco S. Pavone
Part V: ECM Proteins -- Purification, Quantification, and Analysis
28. Methods for Quantifying Fibrillar Collagen Alignment Yuming Liu, Adib Keikhosravi, Guneet S. Mehta, Cole R. Drifka, and Kevin W. Eliceiri
29. Exploring the Nano-Surface of Collagenous and Other Fibrotic Tissues with AFM Andreas Stylianou, Vasiliki Gkretsi, Costas S. Patrickios, and Trianatafyllos Stylianopoulos
30. Spectral Unmixing Methods and Tools for the Detection and Quantitation of Collagen and Other Macromolecules in Tissue Specimens Zachary T. Harmany, Farzad Fereidouni, and Richard M. Levenson
31. Simple Analysis of Deposited Gene Expression Datasets for the Non-Bioinformatician: How to Use GEO for Fibrosis Research Yang Guo, Richard Townsend, and Lam C. Tsoi
This volume describes state-of-the-art protocols that serve as “recipes” for scientists concentrating on fibrosis research. This book is divided into four sections. Part I focuses on animal models of fibrosis and covers topics such as mimicking fibrosis in the lungs, skin, liver and heart, and generating transgenic mouse models. Part II discusses cell culture systems, where the chapters explore cell types important for the development of fibrosis. Part III looks at the purification, quantification, and analysis of the ECM proteins, and Part IV describes computer-assisted methods such as quantifying fibrillar collagen alignment and exploring the nano-surface of collagen with atomic force microscopy (AFM). 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.
Cutting-edge and practical, Fibrosis: Methods and Protocols is a valuable resource aimed at outstanding quality and repeatability of research experiments in the fibrosis field.