Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

Part I: Perspective

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 Rittie, 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

15. Cell-Populated Collagen Lattice Models
Beate Eckes, Fang Wang, Laure Rittie, Gabriele Scherr, and Paola Zigrino

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 Rittie, 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 Rittie

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 Joao Quintas Coentro, Hector Capella-Monsonis, Valeria Graceffa, Zhuning Wu, Anne Maria Mullen, Michael Raghunath, and Dimitrios I. Zeugolis

23. Methods for the Assessment of Active Transforming Growth Factor- 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 Rittie

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