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Full Description
Studies over the past decade have continued to bring tremendous advances to our understanding of bone biology. New pathways have been discovered and expanded our knowledge of the ways in which genes and gene products affect bone cells and thereby bone mass and bone strength. In Bone Research Protocols, Second Edition, expert researchers in the field detail many methods commonly used to study bone biology. Focusing mainly on in vitro methods, this volume gives techniques for isolation, culture and functional analysis of all bone cell types and details a range of imaging methods, including light and ultrastructural microscopy and live cell imaging. Some important in vivo techniques are included, such as analysis of bone resorption and imaging using X rays, fluorescent or luminescent techniques. Methods for study of proteins and nucleic acid are included and methods for analysis of bone composition, measurement of bone strength, and response to mechanical stimulation are described. 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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Bone Research Protocols, Second Edition seeks to aid scientists in the bone field to establish new techniques in their laboratories.
Contents
Primary Human Osteoblast Cultures.- Osteoblast Isolation from Murine Calvaria and Long Bones.- Rat Osteoblast Cultures.- Isolation of primary avian Osteocytes.- Isolation of Mouse Osteocytes using Cell Fractionation for Gene Expression Analysis.- Studying Osteocyte Function using the Cell Lines MLO-Y4 and MLO-A5.- Isolation, Differentiation and Characterisation of Human Bone Marrow Stem Cells in vitro and in vivo.- Rodent Osteoclast Cultures.- Isolation and Culture of Primary Chicken Osteoclasts.- ISOLATION AND PURIFICATION OF RABBIT OSTEOCLASTS.- Generation of Human Osteoclasts from Peripheral Blood.- Osteoclast Formation in Mouse Co-Cultures.- Rankl- Mediated Osteoclast Formation from Murine RAW 264.7 Cells .- Transfection of Osteoclasts and Osteoclast Precursors.- Analysis of Signalling Pathways by Western Blotting and Immunoprecipitation.- Analysis of Transcriptional Regulation in Bone Cells.- Extraction of Nucleic Acids from Bone.- Analysis of Gene Expression in Bone by Quantitative RT/PCR .- Histomorphometry in Rodents.- Studying Gene Expression in Bone by in situ Hybridization.- Immunostaining of Skeletal Tissue.- Techniques for the Study of Apoptosis in Bone.- Transmission Electron Microscopy of Bone.- Scanning Electron Microscopy of Bone.- Fluorescence Imaging of Osteoclasts using Confocal Microscopy.- Live Imaging of Bone Cell and Organ Cultures.- Analysis of Bone Architecture in Rodents using Micro Computed Tomography.- Bone Measurements by Peripheral Quantitative Computed Tomography in Rodents.- Quantitative X-ray Imaging of Rodent Bone by Faxitron.- Bioluminescence Imaging of Bone Metastasis in Rodent.- Fourier Transform Infrared Imaging of Bone.- Raman Microscopy of Bone.- The Calvarial Injection Assay.- Ovariectomy/Orchidectomy in Rodents.- Mechanical Properties of Bone ex vivo.- Mechanical Stimulation of Bone Cells using Fluid Flow.- using CELL and organ culture models TO analyse responses OF bone cellS to mechanical STIMULATION .In vivo Mechanical Loading.
