Bone Research Protocols / Edition 1

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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.

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Editorial Reviews

From the Publisher
"Bone Research Prools 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 prools 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
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Product Details

  • ISBN-13: 9781617373213
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 11/9/2010
  • Series: Methods in Molecular Medicine Series, #80
  • Edition description: Softcover reprint of hardcover 1st ed. 2003
  • Edition number: 1
  • Pages: 450
  • Product dimensions: 5.98 (w) x 9.02 (h) x 0.94 (d)

Table of 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 Immunohishemistry
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


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