Bone Research Protocols

ISBN-13: 9781617373213 / Angielski / Miękka / 2010 / 450 str.

Miep H. Helfrich; Stuart H. Ralston

Opis

The last decade has seen a tremendous advance in our understanding of bone biology. The genes responsible for the majority of rare inherited bone disorders have been identified and much progress has been made in the identification of genes in polygenic disorders such as Paget s disease and complex multigene diseases such as osteoporosis. Transgenic technology has identified further genes, sometimes unexpectedly, with profound effects on bone. This wealth of new genetic information will undoubtedly lead to extensive cell biological studies to understand the mechanisms by which these gene products affect bone mass and bone strength. In Bone Research Protocols a catalogue of protocols has been assembled to perform such mechanistic studies. In the tradition of the Methods in Molecular Medicine series, the chapters are practical laboratory protocols that should enable the reader to carry out the techniques from scratch. To our knowledge this is the first time such a truly practical manual on well-established bone methods has been assembled, and this volume aims to be complementary to and follow on from the more theoretical Methods in Bone Biology, edited by Arnett and Henderson (1)."

Recenzje

"Bone Research Protocols is a most welcome compilation of methods. In it, many of the most productive laboratories in some of the most challenging areas of cell biology reveal detailed protocols that have been used to solve key problems. Short of spending a sabbatical in one of the eminent laboratories that contributed, this volume provides the best possible introduction to these methods." - TRENDS in Endocrinology and Metabolism

Spis treści

Part I. Culture of Osteoblasts and Osteocytes Human Osteoblast Culture James A. Gallagher Osteoblast Isolation from Murine Calvariae and Long Bones Astrid Bakker and Jenneke Klein-Nulend Mineralizing Fibroblast-Colony-Forming Assays Andrew Scutt, Lynsey Reading, Nanette Scutt, and Karen Still Osteocyte Isolation and Culture Peter J. Nijweide, Arie van der Plas, Marcel J. Alblas, and Jenneke Klein-Nulend Part II. Culture of Osteoclasts. Isolated Osteoclast Cultures Astrid Hoebertz and Timothy R. Arnett Primary Isolation and Culture of Chicken Osteoclasts Patricia Collin-Osdoby, Fred Anderson, and Philip Osdoby Isolation and Purification of Rabbit Osteoclasts Fraser P. Coxon, Julie C. Frith, Helena L. Benford, and Michael J. Rogers Generating Human Osteoclasts from Peripheral Blood Afsie Sabokbar and Nicholas A. Athanasou Generating Human Osteoclasts from Bone Marrow and Peripheral Blood Adrienne M. Flanagan and Helen M. Massey Generating Murine Osteoclasts from Bone Marrow Naoyuki Takahashi, Nobuyuki Udagawa, Sakae Tanaka, and Tatsuo Suda Osteoclast Formation in the Mouse Coculture Assay Robert J. van 't Hof RANKL-Mediated Osteoclast Formation from Murine RAW 264.7 Cells Patricia Collin-Osdoby, Xuefeng Yu, Hong Zheng, and Philip Osdoby Part III. Bone Organ Culture Analysis of Osteoclast Function in Mouse Calvarial Cultures Michael J. Marshall and Marit N. Rowlands Assessing Bone Formation Using Mouse Calvarial Organ Culture I. Ross Garrett Part IV. Histological and Ultrastructural Techniques In Situ Hybridization and In Situ Reverse Transcription Polymerase Chain Reaction in Human Bone Sections Andrew P. Mee and Judith A. Hoyland Techniques for the Study of Apoptosis in Bone Brendon S. Noble and Hazel Y. Stevens Protein Localization in Wax-Embedded and Frozen Sections of Bone Using Immunohistochemistry Sharyn Bord Detection of Noncollagenous Bone Proteins in Methylmethacrylate-Embedded Human Bone Sections Johannes P. T. M. van Leeuwen and Pieter Derkx Fluorescence Imaging of Bone-Resorbing Osteoclasts by Confocal Microscopy Stephen A. Nesbitt and Michael A. Horton Bone Histomorphometry Shobna Vedi and Juliet Compston Transmission Electron Microscopy of Bone Vincent Everts, Anneke Niehof, and Wouter Beertsen Scanning Electron Microscopy of Bone Deborah Marshall, Miep H. Helfrich, and Richard M. Aspden Part V. Imaging Techniques Bone Measurement by Peripheral Quantitative Computed Tomography in Rodents Jürg A. Gasser Part VI. In Vivo Techniques. Studies of Local Bone Remodeling: The Calvarial Injection Assay Robert J. van 't Hof, Claire E. Clarkin, and Kenneth J. Armour Inflammation-Induced Osteoporosis: The IMO Model Kenneth J. Armour and Katharine E. Armour Ovariectomy and Estrogen Replacement in Rodents Jade W. M. Chow Part VII. Mechanical Loading Techniques Mechanical Testing of Bone Ex Vivo Richard M. Aspden Bone Cell Responses to Fluid Flow Hazel Y. Stevens and John A. Frangos Methods for Analyzing Bone Cell Responses to Mechanical Loading Using In Vitro Monolayer and Organ Culture Models Andrew A. Pitsillides, Victoria Das-Gupta, Dominic Simon, and Simon C. F. Rawlinson Part VIII. Molecular Analysis Extraction of Nucleic Acids from Bone Tracy L. Stewart and Val Mann Analysis of Gene Expression in Bone by Quantitative RT-PCR Deborah Ireland Index

Blurb

The past decade has witnessed the discovery of many genes with profound effects on bone. In Bone Research Protocols, recognized authorities with extensive hands-on experience present a wide-ranging catalog of laboratory techniques that will help researchers to investigate the mechanisms by which these genes affect bone mass and strength. Described in step-by-step detail, these readily reproducible methods cover such topics as the isolation and culture of bone cells, the preparation of bone tissue for histological and ultrastructural analysis, methods for the measurement of bone strength and for mechanical studies, and how to use digital imaging techniques in the analysis of bone. Though in vitro methods receive central attention, there are also protocols for in vivo analyses of bone formation and resorption.
State-of-the-art and highly practical, Bone Research Protocols offers both beginning and advanced laboratory researchers a major compendium of well-established techniques for studying bone cell function and metabolism.