James A. GallagherAstrid Bakker and Jenneke Klein-NulendAndrew Scutt and Lynsey Reading and Nanette Scutt and Karen StillPeter J. Nijweide and Arie van der Plas and Marcel J. Alblas and Jenneke Klein-NulendAstrid Hoebertz and Timothy R. ArnettPatricia Collin-Osdoby and Fred Anderson and Philip OsdobyFraser P. Coxon and Julie C. Frith and Helena L. Benford and Michael J. RogersAfsie Sabokbar and Nicholas A. AthanasouAdrienne M. Flanagan and Helen M. MasseyNaoyuki Takahashi and Nobuyuki Udagawa and Sakae Tanaka and Tatsuo SudaRobert J. van 't HofPatricia Collin-Osdoby and Xuefeng Yu and Hong Zheng and Philip OsdobyMichael J. Marshall and Marit N. RowlandsI. Ross GarrettAndrew P. Mee and Judith A. HoylandBrendon S. Noble and Hazel Y. StevensSharyn BordJohannes P. T. M. van Leeuwen and Pieter DerkxStephen A. Nesbitt and Michael A. HortonShobna Vedi and Juliet CompstonVincent Everts and Anneke Niehof and Wouter BeertsenDeborah Marshall and Miep H. Helfrich and Richard M. AspdenJurg A. GasserRobert J. van 't Hof and Claire E. Clarkin and Kenneth J. ArmourKenneth J. Armour and Katharine E. ArmourJade W. M. ChowRichard M. AspdenHazel Y. Stevens and John A. FrangosAndrew A. Pitsillides and Victoria Das-Gupta and Dominic Simon and Simon C. F. RawlinsonTracy L. Stewart and Val MannDeborah Ireland

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