By Thomas A. Owen, Lydia C. Pan (auth.), Jennifer J. Westendorf (eds.)
In fresh years, the research of bone cells and tissues on the mobile and molecular degrees in quite a few versions has revolutionized the sphere. In Osteoporosis: equipment and Protocols, top scientists from worldwide proportion their step by step laboratory protocols for learning bone biology. the subjects lined during this quantity comprise in vitro versions, in vivo types applied for drug trying out, tissue engineering and osteoporosis reviews in both gender, cutting-edge molecular options to evaluate unmarried genes or for worldwide genomic research, robust imaging concepts, and lots of extra. As a quantity within the hugely profitable Methods in Molecular Biology™ sequence, every one bankruptcy presents a short advent, a listing of priceless fabrics, and a Notes part detailing pointers on troubleshooting and heading off identified pitfalls.
Comprehensive and state-of-the-art, Osteoporosis: equipment and Protocols is the right advisor for either new and skilled experimentalists trying to research the devastating silent sickness often called osteoporosis.
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Additional info for Osteoporosis: Methods and Protocols
Oursler Antibody Sorting Using MACS 1. Isolate marrow/spleen precursors as described above and omit overnight culture in M-CSF (see Note 12). 2. Pellet cells by centrifugation at 100 g for 10 minutes and resuspend cells in 80 µL running buffer per 1 × 107 cells. 3. Add 20 µL MACS microbeads per 1 × 107 cells (see Note 12) and incubate cells at 4 °C for 20 minutes—do not incubate cells on ice. 4. Pellet cells by centrifugation at 1,000 g for 10 minutes and remove supernatant. 5. Resuspend pelleted cells in 1 mL room temperature running buffer.
This will allow for direct measurement of influences on mature cell functions and/or survival. The caveat with this approach is that these cell preparations will not be as pure, with multiple contaminating cell types that will likely influence responses. 1 Preparation on Dentin or Bone Slices 1. 2 mm. Obtain circles with a paper hole punch. Soak in 70% ethanol overnight to sterilize. 2. Rinse slices repeatedly with sterile water and air-dry in a laminar flow hood. 3. Place slices into 48-well plates and add non-adherent precursor cells as in the above RANKL/M-CSF culture protocol.
The CrossLaps for Culture ELISA from Nordic Biosciences can be used to detect collagen type I fragments generated by resorption of either bone or dentin slices. When paired with assessment of osteoclast numbers, this will provide information on osteoclast activity independent of osteoclast differentiation effects (see Note 15). 2 Artificial Matrix Coating Ceramic biomaterial substrates are now being used by some laboratories in place of biologically derived substrates such as dentin or ivory slices.