Myofibroblasts: Methods and Protocols

ISBN-13: 9781071613849 / Angielski / Miękka / 2021

Hinz, Boris

Opis

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-β 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.

Spis treści

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-β 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-García 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 András 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 Muñoz-Cánoves

 

25. Mouse Models of Skin Fibrosis

            Aleix Rius Rigau, Markus Luber, and Jörg 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 Königshoff