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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 expert researchers in the field detail many methods commonly used to study bone biology.
Focusing mainly on in vitro methods, Bone Research Protocols 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 seeks to aid scientists in the bone field to establish new techniques in their laboratories.
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 Glosman
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. Lowik
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
