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-ß 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.
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-ß 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.
Zusammenfassung
Includes cutting-edge techniques
Provides step-by-step detail essential for reproducible results
Contains key implementation advice from the experts
Inhaltsverzeichnis
50 Years of Myofibroblasts: How the Myofibroblast Concept Evolved.- Myofibroblast Functions in Tissue Repair and Fibrosis: An Introduction.- Myofibroblast Markers and Microscopy Detection Methods in Cell Culture and Histology.- Fibroblast and Myofibroblast Subtypes: Single Cell Sequencing.- Myofibroblast Adhesome Analysis by Mass Spectrometry.- Myofibroblast TGF-ß Activation Measurement In Vitro.- Contraction Measurements Using Three-Dimensional Fibrillar Collagen Gel Lattices.- Techniques to Assess Collagen Synthesis, Deposition, and Cross-Linking In Vitro.- Methods for Studying Myofibroblast Apoptotic Pathways.- Determination of Senescent Myofibroblasts in Precision-Cut Lung Slices.- The Scar-in-a-Jar: In Vitro Fibrosis Model for Anti-Fibrotic Drug Testing.- Why Stress Matters: An Introduction.- Soft Substrate Culture to Mechanically Control Cardiac Myofibroblast Activation.- Quantitative Analysis of Myofibroblast Contraction by Traction Force Microscopy.- Nucleocytoplasmic Shuttlingof the Mechanosensitive Transcription Factors MRTF and YAP/TAZ.- Atomic Force Microscopy for Live-Cell and Hydrogel Measurement.- Method for Investigating Fibroblast Durotaxis.- Decellularized Extracellular Matrix (ECM) as a Model to Study Fibrotic ECM Mechanobiology.- Fibrosis on a Chip for Screening of Anti-Fibrosis Drugs.- Animal and Human Models of Tissue Repair and Fibrosis: An Introduction.- Mouse Models of Lung Fibrosis.- Mouse Models of Kidney Fibrosis.- Mouse Models of Liver Fibrosis.- Mouse Models of Muscle Fibrosis.- Mouse Models of Skin Fibrosis.- Mouse Models of Intestinal Fibrosis.- A Rodent Model of Hypertrophic Scarring: Splinting of Rat Wounds.- Three-Dimensional Model of Hypertrophic Scar Using a Tissue-Engineering Approach.- Methods for the Study of Renal Fibrosis in Human Pluripotent Stem Cell-Derived Kidney Organoids.- Decellularized Human Lung Scaffolds as Complex Three-Dimensional Tissue Culture Models to Study Functional Behavior of Fibroblasts.
