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Full Description
Skeletal Development and Repair: Methods and Protocols is a compilation of a variety of skeletal research protocols utilizing the laboratory mouse as the platform for surgical manipulation and/or transplantation as well as the source of tissues and cells for in vitro culture and analyses. Chapters are written by experts in the field and cover topics including surgical, transplantation and organ culture methods that permit analyses of skeletal tissues undergoing repair in vivo and permits analyses of cellular interactions ex vivo, histological and molecular techniques developed to study gene and protein expression in whole embryos, skeletal tissues and tissue sections and in vitro primary cell culture protocols designed to assay gene function in specific cell populations. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Skeletal Development and Repair: Methods and Protocols is a comprehensive laboratory manual for all levels of basic research scientists working in the broad fields of skeletal development and skeletal repair research.
Contents
Overview of Skeletal Development.- Overview of Skeletal Repair (Fracture Healing and Its Assessment).- Generation of Closed Transverse Fractures in Small Animals.- The Murine Femoral Bone Graft Model and a Semi-Automated Histomorphometric Analysis Tool.- Surgical Induction of Posttraumatic Osteoarthirits.- A Mouse Model of Flexor Tendon Repair.- Cartilage Explant Cultures.- Renal Capsule Transplantations to Assay Skeletal Angiogenesis.- Whole Mount Skeletal Staining.- Tips and Techniques for Processing and Sectioning Undecalcified Murine Bone Specimens.- Preparation of Thin Sections from Non-fixed and Undecalcified Hard Tissues Using Kawamoto's Film Method (2012).- Demineralized Murine Skeletal Histology.- Beta-Galactosidase Staining in the Skeleton.- Whole-Mount In Situ Hybridization on Murine Skeletogenic Tissues.- Non-Radioactive In Situ Hybridization on Skeletal Tissue Sections.- Radioactive In Situ Hybridization to Detect Gene Expression in Skeletal Tissue Sections.- Proliferation Assays (BrdU and EdU) on Skeletal Tissue Sections.- TUNEL Assay on Skeletal Tissue Sections to Detect Apoptosis.- Analysis of Chondrogenesis Using Micromass Cultures of Limb Mesenchyme.- Isolation and Culture of Murine Primary Chondrocytes.- Bone Marrow Stromal Cell Assays - In Vitro and In Vivo.- Isolation and Culture of Neonatal Mouse Calvarial Osteoblasts.- RANKL-BasedOsteoclastogenic Assays from Murine Bone Marrow Cells.- Hematopoietic Stem Cell Cultures and Assays.
