Bone research protocols / edited by Miep H. Helfrich, Stuart H. Ralston.

Format:

Book

Contributor:

Helfrich, Miep H.

Ralston, Stuart.

Series:

Methods in molecular medicine ; 80.

Methods in molecular medicine ; 80

Language:

English

Subjects (All):

Bones--Cultures and culture media--Laboratory manuals.

Bones.

Bone cells--Laboratory manuals.

Bone cells.

Bone and Bones--cytology.

Cell Culture Techniques.

Histological Techniques.

Osteoblasts.

Osteoclasts.

Medical Subjects:

Bone and Bones--cytology.

Cell Culture Techniques.

Histological Techniques.

Osteoblasts.

Osteoclasts.

Genre:

Laboratory manuals.

Physical Description:

xiv, 448 pages : illustrations ; 23 cm.

Place of Publication:

Totowa, N.J. : Humana Press, [2003]

Summary:

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.

Contents:

Part I Culture of Osteoblasts and Osteocytes

1 Human Osteoblast Culture / James A. Gallagher 3

2 Osteoblast Isolation from Murine Calvariae and Long Bones / Astrid Bakker, Jenneke Klein-Nulend 19

3 Mineralizing Fibroblast-Colony-Forming Assays / Andrew Scutt, Lynsey Reading, Nanette Scutt, Karen Still 29

4 Osteocyte Isolation and Culture / Peter J. Nijweide, Arie van der Plas, Marcel J. Alblas, Jenneke Klein-Nulend 41

Part II Culture of Osteoclasts

5 Isolated Osteoclast Cultures / Astrid Hoebertz, Timothy R. Arnett 53

6 Primary Isolation and Culture of Chicken Osteoclasts / Patricia Collin-Osdoby, Fred Anderson, Philip Osdoby 65

7 Isolation and Purification of Rabbit Osteoclasts / Fraser P. Coxon, Julie C. Frith, Helena L. Benford, Michael J. Rogers 89

8 Generating Human Osteoclasts from Peripheral Blood / Afsie Sabokbar, Nicholas A. Athanasou 101

9 Generating Human Osteoclasts from Bone Marrow and Peripheral Blood / Adrienne M. Flanagan, Helen M. Massey 113

10 Generating Murine Osteoclasts from Bone Marrow / Naoyuki Takahashi, Nobuyuki Udagawa, Sakae Tanaka, Tatsuo Suda 129

11 Osteoclast Formation in the Mouse Coculture Assay / Robert J. van 't Hof 145

12 RANKL-Mediated Osteoclast Formation from Murine RAW 264.7 Cells / Patricia Collin-Osdoby, Xuefeng Yu, Hong Zheng, Philip Osdoby 153

Part III Bone Organ Culture

13 Analysis of Osteoclast Function in Mouse Calvarial Cultures / Michael J. Marshall, Marit N. Rowlands 169

14 Assessing Bone Formation Using Mouse Calvarial Organ Culture / I. Ross Garrett 183

Part IV Histological and Ultrastructural Techniques

15 In Situ Hybridization and In Situ Reverse Transcription Polymerase Chain Reaction in Human Bone Sections / Andrew P. Mee, Judith A. Hoyland 201

16 Techniques for the Study of Apoptosis in Bone / Brendon S. Noble, Hazel Y. Stevens 225

17 Protein Localization in Wax-Embedded and Frozen Sections of Bone Using Immunohistochemistry / Sharyn Bord 237

18 Detection of Noncollagenous Bone Proteins in Methylmethacrylate-Embedded Human Bone Sections / Johannes P. T. M. van Leeuwen, Pieter Derkx 249

19 Fluorescence Imaging of Bone-Resorbing Osteoclasts by Confocal Microscopy / Stephen A. Nesbitt, Michael A. Horton 259

20 Bone Histomorphometry / Shobna Vedi, Juliet Compston 283

21 Transmission Electron Microscopy of Bone / Vincent Everts, Anneke Niehof, Wouter Beertsen 299

22 Scanning Electron Microscopy of Bone / Deborah Marshall, Miep H. Helfrich, Richard M. Aspden 311

Part V Imaging Techniques

23 Bone Measurements by Peripheral Quantitative Computed Tomography in Rodents / Jurg A. Gasser 323

Part VI In Vivo Techniques

24 Studies of Local Bone Remodeling: The Calvarial Injection Assay / Robert J. van 't Hof, Claire E. Clarkin, Kenneth J. Armour 345

25 Inflammation-Induced Osteoporosis: The IMO Model / Kenneth J. Armour, Katharine E. Armour 353

26 Ovariectomy and Estrogen Replacement in Rodents / Jade W. M. Chow 361

Part VII Mechanical Loading Techniques

27 Mechanical Testing of Bone Ex Vivo / Richard M. Aspden 369

28 Bone Cell Responses to Fluid Flow / Hazel Y. Stevens, John A. Frangos 381

29 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, Simon C. F. Rawlinson 399

Part VIII Molecular Analysis

30 Extraction of Nucleic Acids from Bone / Tracy L. Stewart, Val Mann 425

31 Analysis of Gene Expression in Bone by Quantitative RT-PCR / Deborah Ireland 433.

Notes:

Includes bibliographical references and index.

ISBN:

1588290441

OCLC:

51271023