Skeletal Development and Repair

This second edition provides a comprehensive laboratory manual on skeletal development and skeletal repair research utilizing mouse models. Chapters provide methods and protocols on the most current and cutting-edge techniques in skeletal development and repair, histological, cellular, and molecular techniques. 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. Authoritative and cutting-edge, Skeletal Development and Repair: Methods and Protocols-Second Edition aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt
Overview of Skeletal Development.- Overview of Skeletal Repair (Fracture Healing and Its Assessment).- Advantages and Limitations of Cre Mouse Lines Used in Skeletal Research.- Generation of Closed Transverse Fractures in Small Animals.- A Mouse Femoral Ostectomy Model to Assess Bone Graft Substitutes.- Surgical Induction of Posttraumatic Osteoarthritis in the Mouse.- Parabiosis: Assessing the Effects of Circulating Cells and Factors on the Skeleton.- Murine Limb Bud Organ Cultures for Studying Musculoskeletal Development.- Murine Limb Explant Cultures to Assess Cartilage Development.- Renal Capsule Transplantation to Assay Angiogenesis in Skeletal Development and Repair.- MicroCT for Scanning and Analysis of Mouse Bones.- Four-Point Bending Testing for Mechanical Assessment of Mouse Bone Structural Properties.- Phenotyping Intact Mouse Bones using Bone CLARITY.- Processing and Sectioning Undecalcified Murine Bone Specimens.- Preparation of Thin Frozen Sections from Nonfixed And Undecalcified Hard Tissues Using Kawamoto's Film Method (2020).- Demineralized Murine Skeletal Histology.- Assessment of Osteocytes: Techniques for Studying Morphological and Molecular Changes Associated With Perilacunar/ Canalicular Remodeling Of the Bone Matrix.- Cell Lineage Tracing: Co-localization of Cell Lineage Markers with a Fluorescent Reporter.- Immunofluorescent Staining of Adult Murine Paraffin-embedded Skeletal Tissue.- Detection of Hypoxic Regions in the Bone Microenvironment.- EdU-based Assay of Cell Proliferation and Stem Cell Quiescence in Skeletal Tissue Sections.- Whole-mount In Situ Hybridization in Murine Tissues.- Bone Marrow Stromal Cell Assays In Vitro and In Vivo.- Isolation and Culture of Periosteum Derived Progenitor Cells from Mice.- Isolation and Culture of Murine Primary Chondrocytes: Costal and Growth Plate Cartilage.- Isolation and Culture of Neonatal Mouse Calvarial Osteoblasts.-Mitochondrial Function and Metabolism of Cultured Skeletal Cells.- Radiolabeled Amino Acid Uptake Assays in Primary Bone Cells and Bone Explants.- RANKL-based Osteoclastogenic Assay from Murine Bone Marrow Cells.- Hematopoietic Stem Cell Cultures and Assays.