Osteoporosis: Methods and Protocols by Thomas A. Owen, Lydia C. Pan (auth.), Jennifer J. Westendorf

By Thomas A. Owen, Lydia C. Pan (auth.), Jennifer J. Westendorf (eds.)

In fresh years, the learn of bone cells and tissues on the mobile and molecular degrees in various types has revolutionized the sphere. In Osteoporosis: tools and Protocols, prime scientists from all over the world percentage their step by step laboratory protocols for learning bone biology. the subjects lined during this quantity contain in vitro versions, in vivo types applied for drug trying out, tissue engineering and osteoporosis experiences in both gender, state of the art molecular innovations to evaluate unmarried genes or for worldwide genomic research, robust imaging options, and lots of extra. As a quantity within the hugely winning Methods in Molecular Biology™ sequence, each one bankruptcy offers a short creation, a listing of priceless fabrics, and a Notes part detailing pointers on troubleshooting and warding off recognized pitfalls.

Comprehensive and state-of-the-art, Osteoporosis: tools and Protocols is the suitable consultant for either new and skilled experimentalists trying to examine the devastating silent ailment referred to as osteoporosis.

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4. Place the mouse pups under the hood. 5. Pick the first pup with the large forceps. 6.

11. Mouse calvarial cultures show a marked loss of differentiation capacity with subculturing; therefore, it is advisable to use primary cultures if osteoblastic differentiation is the desired endpoint. In contrast, rat calvarial cells can be expanded for three to four population doublings without significant loss of the potential for mineralized nodule formation (13). 12. For experiments with osteoblastic differentiation as an endpoint, plating calvarial osteoblasts in wells <2 cm2 (24-well plate) or rodent bone marrow cells in wells <9 cm2 (6-well) will result in an uneven distribution of the osteoblastic precursor cells among the wells.

4. 5. Sterile PBS. 100-mm tissue culture dishes. Staining buffer: Sterile PBS supplemented with 1% BSA. PE (or another suitable fluorophore)-conjugated CD11b or CD14 antibody. Phenol red-free αMEM with L-glutamine and sodium pyruvate supplemented with 10% FBS and 1% antimycotic/antibiotic solution. 6. 0 µg/mL in water, stored in aliquots at −80 °C until diluted (100×) in culture medium. 7. 1% BSA stored in aliquots at −20 °C until diluted (100×) in culture medium. 2 1. 2. 3. 4. 5. 6. 7. 8. 9. 10.

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