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Bone Research Protocols
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Main description:

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).


Contents:

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
Jurg 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


PRODUCT DETAILS

ISBN-13: 9781617373213
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 462
Weight: 782g
Availability: Available
Subcategories: Orthopaedics and Fractures
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