Bone research protocols
Author(s)
Bibliographic Information
Bone research protocols
（Methods in molecular biology / John M. Walker, series editor, 816）
Humana Press, c2012
2nd ed
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Note
Includes bibliographical references and index
Description and Table of Contents
Description
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 (TM) 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.
Table of Contents
Part I: Culture of Osteoblasts and Osteocytes.- 1. Primary Human Osteoblast Cultures.- Jane P. Dillon, Victoria J. Waring-Green, Adam M. Taylor, Peter Wilson, Mark Birch, Alison Gartland, James A. Gallagher .- 2. Osteoblast Isolation from Murine Calvaria and Long Bones.- Jenneke Klein-Nulend and Astrid D. Bakker .- 3. Rat Osteoblast Cultures.- Isabel R. Orriss, Sarah E.B. Taylor and Timothy R. Arnett.- 4. Isolation of Primary Avian Osteocytes.- Cor M. Semeins, Astrid D. Bakker, Jenneke Klein-Nulend.- 5. Isolation of Mouse Osteocytes using Cell Fractionation for Gene Expression Analysis. .- Christine Halleux, Ina Kramer, Cyril Allard and Michaela Kneissel .- 6. Studying Osteocyte Function using the Cell Lines MLO-Y4 and MLO-A5.- Jennifer Rosser and Lynda F. Bonewald.- 7. Isolation, Differentiation and Characterisation of Human Bone Marrow Stem Cells in vitro and in vivo.- Rahul S. Tare, Peter D. Mitchell, Janos Kanczler, Richard O.C. Oreffo.- Part II: Culture of Osteoclasts.- 8. Rodent Osteoclast Cultures.- Isabel R. Orris and Timothy R. Arnett.- 9. Isolation and Culture of Primary Chicken Osteoclasts.- Patricia Collin-Osdoby and Phillip Osdoby.- 10. Isolation and Purification of Rabbit Osteoclasts .- Fraser P. Coxon, Michael J. Rogers and Julie C. Crockett.- 11. Generation of Human Osteoclasts from Peripheral Blood.- Kim Henriksen, Morten A. Karsdal, Adam Taylor, Denise Tosh and Fraser P. Coxon.- 12. Osteoclast Formation in Mouse Co-Cultures.- Cecile Itzstein and Robert J. van't Hof.- 13. Rankl-Mediated Osteoclast Formation From Murine RAW 264.7 Cells.- Patricia Collin-Osdoby and Phillip Osdoby.- Part III: Biochemical and Molecular Analysis of Bone Cells.- 14. Transfection of Osteoclasts and Osteoclast Precursors.- Julie C. Crockett, David J. Mellis and Adam Taylor.- 15. Analysis of Signalling Pathways by Western Blotting and Immunoprecipitation.- Aymen I. Idris.- 16. Analysis of Transcriptional Regulation in Bone Cells .- Huilin Jin and Stuart Ralston.- 17. Extraction of Nucleic Acids from Bone.- Alun Hughes, Tracey L. Steward and Val Mann.- 18. Analysis of Gene Expression in Bone by Quantitative RT/PCR.- Alun Hughes.- Part IV: Microscopial Techniques.- 19. Histomorphometry in Rodents .- Reinhold G. Erben and Martin Gloesman.- 20. Studying Gene Expression in Bone by in situ Hybridization .- Ina Kramer, Rishard Sali, Mira Susa, and Michaela Kneissel.- 21. Immunostaining of Skeletal Tissue .- Tobias Kurth and Cosimo De Bari.- 22. Techniques for the Study of Apoptosis in Bone.- Sudeh Riahi and Brendon Noble .- 23. Transmission Electron Microscopy of Bone.- Vincent Everts, Anneke Niehof, Wikky Tigchelarr-Gutter and Wouter Beertsen.- 24. Scanning Electron Microscopy of Bone.- Alan Boyde.- 25. Fluorescence Imaging of Osteoclasts using Confocal Microscopy.- Fraser P. Coxon.- 26. Live Imaging of Bone Cell and Organ Cultures .- Sarah L. Dallas and Pat A. Veno.- Part V: Imaging TEchniques .- 27. Analysis of Bone Architecture in Rodents using Micro Computed Tomography .- Rob van't Hof.- 28. Bone Measurments by Peripheral Quantitatvie Computer Tomography in Rodents .- Jurg Gasser, and Johannes Willnecker.- 29. Quantitative X-Ray Imaging of Rodent Bone by Faxitron.- JH Duncan Basset, Anne van der Spek, Apostolos Gogakos, Graham R Williams.- 30. Bioluminescence Imaging of Bone Metastasis in Rodents .- Thomas J.A. Snoeks, Ermond van Beek, Ivo Que, Eric L. Kaijzel and Clemens W.G.M. Loewik.- 31. Fourier Transform Infrared Imaging of Bone .- Elefterios P. Paschalis.- 32. Raman Microscopy of Bone.- Simon R. Goodyear and Richard M. Aspden.- Part VI: In Vivo Techniques.- 33. The Calvarial Injection Assay.- Robert J. van't Hof.- 34. Ovariectomy/Orchidectomy in Rodents.- Aymen I. Idris.- Part VIII: Mechanical Loading Techniques.- 35. Mechanical Properties of Bone Ex Vivo .- Simon R. Goodyear and Rchiard M. Aspden.- 36. Mechanical Stimulation of Bone Cells using Fluid Flow.- Carmen Huesa and Astrid D. Bakker.- 37. Using Cell and Organ Culture Models to Analyse Responsed of Bone Cells to Mechanical Stimulation.- Andres A. Pitsillides and Simon C. F. Rawlinson.- 38. In Vivo Mechanical Loading .- Roberto Lopes de Souza and Leanne Saxon.
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