By Miep H. Helfrich, Stuart H. Ralston
The decade has obvious a major strengthen in our figuring out of bone biology. The genes liable for nearly all of infrequent inherited bone problems were pointed out and masses development has been made within the identity of genes in polygenic issues corresponding to Paget’s ailment and intricate multigene ailments comparable to osteoporosis. Transgenic expertise has pointed out extra genes, occasionally abruptly, with profound results on bone. This wealth of latest genetic info will absolutely bring about wide cellphone organic reviews to appreciate the mechanisms during which those gene items impact bone mass and bone power. In Bone examine Protocols a listing of protocols has been assembled to accomplish such mechanistic stories. within the culture of the equipment in Molecular drugs sequence, the chapters are useful laboratory protocols that are meant to permit the reader to hold out the concepts from scratch. To our wisdom this is often the 1st time this sort of really sensible guide on well-established bone equipment has been assembled, and this quantity goals to be complementary to and persist with on from the extra theoretical equipment in Bone Biology, edited by way of Arnett and Henderson (1).
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Additional resources for Bone Research Protocols (Methods in Molecular Medicine)
The isolation of these cells is facilitated by mild EDTA treatments alternating with collagenase treatments. The choice of fetal chicken calvariae is important because in this species the periostea from both calvaria surfaces can be dissected relatively easily, which results in the absence of most periosteal cells in the collagenase released cell population. Furthermore, fetal chicken calvariae have a much broader layer of osteoid on their surfaces than mouse or rat calvariae. Finally, the availability of a monoclonal antibody (MAb) that specifically recognizes osteocytes allows the purification of osteocytes from mixed cell populations (immunodissection).
Endocrinology 119, 1776–1785. 12. Fedarko, N. , and Robey, P. G. (1992) Age-related changes in hyaluronan, proteoglycan, collagen and osteonectin synthesis by human bone cells. J. Cell Physiol. 151, 215–227. 13. Beresford, J. , et al. (1984) The effects of monocyte-conditioned medium and interleukin 1 on the synthesis of collagenous and non-collagenous proteins by mouse bone and human bone cells in vitro. Biochim. Biophysica. Acta Gen. Subj. 801, 58–65. 14. , Wood, D. , and Russell, R. G. (1985) Stimulation of the proliferation of human bone cells in vitro by human monocyte products with interleukin-1 activity in J.
Cells are obtained from the long bones and the calvariae of adult (age 9 wk or older) mice, or the calvariae from neonatal mice pups (age 3–4 d). 2. Instruments All of the following materials have to be sterile. 1. Polystyrene plate and needles for fixing the mice. 2. Scalpels (no. 10 and 11), scissors, tweezers, and curved forceps. 3. 2-µm disposable filter units. 02/Nulend/19-28/F1 20 2/26/03, 10:44 AM Bone Cells from Calvariae and Long Bones 21 4. 25-cm2 tissue culture flasks (Nunc), six-well tissue plates (Costar), 94/16-mm cellstar Petri dishes (Greiner), and 145/20-mm cellstar (large) Petri dishes (Greiner).