Bone Research Protocols / Edition 1

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Overview

A collection of the latest laboratory techniques for the study of bone and bone tissue. 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.

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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: 9781588290441
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 4/28/2003
  • Series: Methods in Molecular Medicine Series
  • Edition number: 1
  • Pages: 468
  • Product dimensions: 6.20 (w) x 9.20 (h) x 1.10 (d)

Table of Contents

Preface
Contributors
1 Human Osteoblast Culture 3
2 Osteoblast Isolation from Murine Calvariae and Long Bones 19
3 Mineralizing Fibroblast-Colony-Forming Assays 29
4 Osteocyte Isolation and Culture 41
5 Isolated Osteoclast Cultures 53
6 Primary Isolation and Culture of Chicken Osteoclasts 65
7 Isolation and Purification of Rabbit Osteoclasts 89
8 Generating Human Osteoclasts from Peripheral Blood 101
9 Generating Human Osteoclasts from Bone Marrow and Peripheral Blood 113
10 Generating Murine Osteoclasts from Bone Marrow 129
11 Osteoclast Formation in the Mouse Coculture Assay 145
12 RANKL-Mediated Osteoclast Formation from Murine RAW 264.7 Cells 153
13 Analysis of Osteoclast Function in Mouse Calvarial Cultures 169
14 Assessing Bone Formation Using Mouse Calvarial Organ Culture 183
15 In Situ Hybridization and In Situ Reverse Transcription Polymerase Chain Reaction in Human Bone Sections 201
16 Techniques for the Study of Apoptosis in Bone 225
17 Protein Localization in Wax-Embedded and Frozen Sections of Bone Using Immunohistochemistry 237
18 Detection of Noncollagenous Bone Proteins in Methylmetacrylate-Embedded Human Bone Sections 249
19 Fluorescence Imaging of Bone Resorbing Osteoclasts by Confocal Microscopy 259
20 Bone Histomorphometry 283
21 Transmission Electron Microscopy of Bone 299
22 Scanning Electron Microscopy of Bone 311
23 Bone Measurements by Peripheral Quantitative Computed Tomography in Rodents 323
24 Studies of Local Bone Remodeling: The Calvarial Injection Assay 345
25 Inflammation-Induced Osteoporosis: The IMO Model 353
26 Ovaridectomy and Estrogen Replacement in Rodents 361
27 Mechanical Testing of Bone Ex Vivo 369
28 Bone Cell Responses to Fluid Flow 381
29 Methods for Analyzing Bone Cell Responses to Mechanical Loading Using In Vitro Monolayer and Organ Culture Models 399
30 Extraction of Nucleic Acids from Bone 425
31 Analysis of Gene Expression in Bone by Quantitative RT-PCR 433
Index 441
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