Bone Research Protocols by Miep H. Helfrich, Stuart H. Ralston

By Miep H. Helfrich, Stuart H. Ralston

Laboratory ideas provided right here can assist researchers investigating the mechanisms wherein convinced genes have an effect on bone mass and power. those reproducible equipment disguise such themes because the isolation and tradition of bone cells, the coaching of bone tissue for histological checking out, and using electronic imaging ideas within the research of bone. even though in vitro equipment obtain such a lot cognizance, there also are protocols for in vivo analyses. The editors are affiliated with the collage of Aberdeen clinical college, united kingdom.

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The condyle of the femur can be grasped and torn off in a similar manner. However, not all of the muscle will come off cleanly and will have to be removed carefully using either scissors or a scalpel. The bone can then be cut as close to the femoral neck as possible using a rotary saw. 3. Recover the bone marrow cells using the method of Dobson et al. 5 mL microfuge tubes or hypodermic needle casings (Fig. 2). (see Note 5). 4. Centrifuge the tubes briefly at 900 g for 5 sec in a microfuge. The marrow cells will pellet at the bottom of the tube.

J. Bone Miner. Res. 13, 1555–1568. 13. Aarden, E. , Wassenaar, A. , Alblas, M. , and Nijweide, P. J. (1996) Immunocytochemical demonstration of extracellular matrix proteins in isolated osteocytes. Histochem. Cell Biol. 106, 495–501. 14. Nakamura, H. and Ozawa, H. (1996) Immunolocalization of CD44 and the ERM family in bone cells of mouse tibiae. J. Bone Miner. Res. 11, 1715–1722. 04/Nijweide/41-50/F1 50 2/26/03, 10:45 AM Isolated Osteoclast Cultures 53 5 Isolated Osteoclast Cultures Astrid Hoebertz and Timothy R.

3. Allow to dry at room temperature. 4. Photograph the dishes to document the amount of mineralization. 5. Destain the dishes by gently shaking with 5% percholoric acid for 5 min. 6. Wash thoroughly with tap water. 5. Collagen 1. 2. 3. 4. 5. 6. Add 5 mL of collagen staining solution to each dish and incubate for 18 h. Wash the dishes under running tap water until the excess dye has washed off. Allow to air-dry at room temperature. Photograph the dishes to document the amount of staining. Destain the dishes by gently shaking with collagen destaining solution.

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