Bone Research Protocols

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Miep H. Helfrich, Stuart H. Ralston
Springer Science & Business Media, 1 feb 2008 - 450 pagine
The last decade has seen a tremendous advance in our understanding of bone biology. The genes responsible for the majority of rare inherited bone disorders have been identified and much progress has been made in the identification of genes in polygenic disorders such as Paget’s disease and complex multigene diseases such as osteoporosis. Transgenic technology has identified further genes, sometimes unexpectedly, with profound effects on bone. This wealth of new genetic information will undoubtedly lead to extensive cell biological studies to understand the mechanisms by which these gene products affect bone mass and bone strength. In Bone Research Protocols a catalogue of protocols has been assembled to perform such mechanistic studies. In the tradition of the Methods in Molecular Medicine series, the chapters are practical laboratory protocols that should enable the reader to carry out the techniques from scratch. To our knowledge this is the first time such a truly practical manual on well-established bone methods has been assembled, and this volume aims to be complementary to and follow on from the more theoretical Methods in Bone Biology, edited by Arnett and Henderson (1).

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Human Osteoblast Culture
3
Osteoblast Isolation from Murine Calvariae and Long Bones
19
and Karen Still 4 Osteocyte Isolation and Culture
29
CULTURE OF OSTEOCLASTS
63
Isolation and Purification of Rabbit Osteoclasts
89
Generating Human Osteoclasts from Peripheral Blood
101
Flanagan and Helen M Massey 10 Generating Murine Osteoclasts from Bone Marrow
113
Naoyuki Takahashi Nobuyuki Udagawa Sakae Tanaka
129
Detection of Noncollagenous Bone Proteins
249
Fluorescence Imaging of BoneResorbing Osteoclasts
259
Bone Histomorphometry
283
Transmission Electron Microscopy of Bone
299
Scanning Electron Microscopy of Bone
311
Bone Measurements by Peripheral Quantitative
323
The IMO Model
353
Ovariectomy and Estrogen Replacement in Rodents
361

Osteoclast Formation in the Mouse Coculture Assay
145
from Murine RAW 264 7 Cells
153
Analysis of Osteoclast Function in Mouse Calvarial Cultures
169
Assessing Bone Formation
183
In Situ Hybridization and In Situ Reverse Transcription
201
Brendon S Noble and Hazel Y Stevens 17 Protein Localization in WaxEmbedded and Frozen Sections
225
of Bone Using Immunohistochemistry
237
Mechanical Testing of Bone Ex Vivo
369
Hazel Y Stevens and John A Frangos 29 Methods for Analyzing Bone Cell Responses to Mechanical
381
Loading Using In Vitro Monolayer and Organ Culture Models
399
Extraction of Nucleic Acids from Bone
425
Analysis of Gene Expression in Bone by Quantitative RTPCR
433
Index
441
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