Description
This detailed book explores the fundamentals of myofibroblast biology in tissue repair, fibrosis, and tumors as well as providing cutting-edge laboratory methods used to investigate myofibroblast functions in physiological and pathological settings in vitro and in vivo, as written by leading academic scientists. Section I of this volume focuses on fundamental methods to study myofibroblast biology and covers topics such as methods for detecting myofibroblasts and senescent myofibroblasts in cell culture and histology, single cell RNA sequencing to identify myofibroblast subsets in fibrotic tissues, and functional assays to assess TGF- activation, myofibroblast apoptosis, or matrix deposition and crosslinking. Section II discusses methods to investigate the mechanobiology of myofibroblasts in vitro, including the fabrication of functional hydrogels with tunable stiffness, the use of atomic force microscopy to characterize matrix and cellular stiffness, as well as molecular assays to assess fibroblast mechanotransduction pathways and durotaxis. Section III describes multiple animal models to investigate myofibroblast functions across organs in vivo as well as human organoid systems, precision tissue slices and decellularized 3D tissue scaffolds to assess myofibroblast functions in relevant human ex vivo models. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and extensive, Myofibroblasts: Methods and Protocols is an essential collection for researchers delving into the processes and effects of these important cells. 1. 50 Years of Myofibroblasts: How the Myofibroblast Concept Evolved Giulio Gabbiani Section I: Fundamental Methods to Study Myofibroblast Biology 2. Myofibroblast Functions in Tissue Repair and Fibrosis: An Introduction Victor J. Thannickal 3. Myofibroblast Markers and Microscopy Detection Methods in Cell Culture and Histology Fereshteh Younesi, Dong Ok Son, Joao Firmino, and Boris Hinz 4. Fibroblast and Myofibroblast Subtypes: Single Cell Sequencing Hesham Soliman, Lin Wei Tung, and Fabio M.V. Rossi 5. Myofibroblast Adhesome Analysis by Mass Spectrometry Christopher A. McCulloch 6. Myofibroblast TGF- Activation Measurement In Vitro Joanne Porte, Gisli Jenkins, and Amanda L. Tatler 7. Contraction Measurements Using Three-Dimensional Fibrillar Collagen Gel Lattices James Griffith and William L. Berry 8. Techniques to Assess Collagen Synthesis, Deposition, and Cross-Linking In Vitro Tamara Rosell-Garca and Fernando Rodriguez-Pascual 9. Methods for Studying Myofibroblast Apoptotic Pathways Yan Zhou and David Lagares 10. Determination of Senescent Myofibroblasts in Precision-Cut Lung Slices Tamara Cruz, Ana L. Mora, and Mauricio Rojas 11. The Scar-in-a-Jar: In Vitro Fibrosis Model for Anti-Fibrotic Drug Testing Simon Stebler and Michael Raghunath Section II: Myofibroblast Mechanobiology 12. Why Stress Matters: An Introduction Daniel J. Tschumperlin 13. Soft Substrate Culture to Mechanically Control Cardiac Myofibroblast Activation Natalie M. Landry, Sunil G. Rattan, and Ian M.C. Dixon 14. Quantitative Analysis of Myofibroblast Contraction by Traction Force Microscopy Shuying Yang, Fernando R. Valencia, Benedikt Sabass, and Sergey V. Plotnikov 15. Nucleocytoplasmic Shuttling of the Mechanosensitive Transcription Factors MRTF and YAP/TAZ Michael Kofler and Andrs Kapus 16. Atomic Force Microscopy for Live-Cell and Hydrogel Measurement Alexander Whitehead, Natalie J. Kirkland, and Adam J. Engler 17. Method for Investigating Fibroblast Durotaxis Hossam Kadry, David Lagares, and Taslim A. Al-Hilal 18. Decellularized Extracellular Matrix (ECM) as a Model to Study Fibrotic ECM Mechanobiology Chiuan-Ren Yeh, Grace C. Bingham, Jagathpala Shetty, Ping Hu, and Thomas H. Barker 19. Fibrosis on a Chip for Screening of Anti-Fibrosis Drugs Mohammadnabi Asmani and Ruogang Zhao Section III: Animal and Human Models to Study Myofibroblast Biology 20. Animal and Human Models of Tissue Repair and Fibrosis: An Introduction David Lagares and Boris Hinz 21. Mouse Models of Lung Fibrosis Olivia Mekhael, Safaa Naiel, Megan Vierhout, Aaron Hayat, Spencer Revill, Soumeya Abed, Mark Inman, Martin Kolb, and Kjetil Ask 22. Mouse Models of Kidney Fibrosis Rafael Kramann and Sylvia Menzel 23. Mouse Models of Liver Fibrosis Aashreya Ravichandra and Robert F. Schwabe 24. Mouse Models of Muscle Fibrosis Antonio L. Serrano and Pura Muoz-Cnoves 25. Mouse Models of Skin Fibrosis Aleix Rius Rigau, Markus Luber, and Jrg H.W. Distler 26. Mouse Models of Intestinal Fibrosis Jiannan Li, Dina Dejanovic, Megan T. Zangara, Jyotsna Chandra, Christine McDonald, and Florian Rieder 27. A Rodent Model of Hypertrophic Scarring: Splinting of Rat Wounds Dong Ok Son and Boris Hinz 28. Three-Dimensional Model of Hypertrophic Scar Using a Tissue-Engineering Approach Veronique J. Moulin 29. Methods for the Study of Renal Fibrosis in Human Pluripotent Stem Cell-Derived Kidney Organoids Alejandro Moran-Horowich and Dario R. Lemos 30. Decellularized Human Lung Scaffolds as Complex Three-Dimensional Tissue Culture Models to Study Functional Behavior of Fibroblasts Gerald Burgstaller, Michael Gerckens, Oliver Eickelberg, and Melanie Knigshoff
