Bone Research ProtocolsMiep 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). |
Sommario
3 | |
19 | |
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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