Abstract
Primary cells are often desirable over clonal cell lines, because they are more likely to retain the presence and activity of certain enzymes and proteins that are often lost in clones. In addition, primary cells more closely resemble the cell in the actual animal than clones do. An often-cited drawback of primary cells is the heterogeneity inherent in them. Primary osteoblast cultures, for example, consist of preosteoblasts, osteoblasts, and osteocytes (if the culture is old enough), and possibly even a few fibroblasts Such a mix of osteoblastic cells, however, could be deemed beneficial and more realistic. The most detrimental cell contaminant is the fibroblast. Fibroblasts, if present in the culture, will eventually take over. We have never had this problem with our cultures Osteoblastic cultures isolated by the method described in this chapter have been shown to be osteoblastic in nature by exhibiting extensive alkaline phosphatase activity (1–3), by forming nodules that stain positive for mineral by a Von Kossa stain (1–5, and by showing a characteristic increase (three-fold) in intracellular cAMP in response to PTH stimulation (2.7×10−8 M for 15 mm) (1).
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Reich, K. M., Gay, C. V., and Frangos, J. A. (1990) Fluid shear stress as a mediator of osteoblast cyclic adenosine monophosphate production. J. Cell Physiol. 143, 100–104.
Hillsley, M. V. (1990) The effects of fluid shear stress and 1,25-dihydroxy-vitamin D3 on collagen and osteocalcin production by osteoblasts. MS Thesis. Pennsylvania State University.
Hillsley, M. V. and Frangos, J. A. (1997) Alkaline phosphatase in osteoblasts is down-regulated by pulsatile fluid flow. Calc. Tissue Int., 60(1), 48–53.
Hillsley, M. V. and Frangos, J. A. (1994) Bone tissue engineering: the role of interstitial fluid flow. Biotechnol. Bioeng. 43, 573–581 (review).
Hillsley, M. V. and Frangos, J. A. (1996) Osteoblast hydraulic conductivity is regulated by calcitonin and parathyroid hormone. J. Bone Miner. Res. 11, 114–124.
Ecarot-Charrier, B., Glorieux, H., van der Rest, M., and Pereira, G. (1983) Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. J. Cell Biol. 96, 639–643.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Hillsley, M.V. (1999). Methods to Isolate, Culture, and Study Osteoblasts. In: Morgan, J.R., Yarmush, M.L. (eds) Tissue Engineering Methods and Protocols. Methods in Molecular Medicine™, vol 18. Humana Press. https://doi.org/10.1385/0-89603-516-6:293
Download citation
DOI: https://doi.org/10.1385/0-89603-516-6:293
Publisher Name: Humana Press
Print ISBN: 978-0-89603-516-4
Online ISBN: 978-1-59259-602-7
eBook Packages: Springer Protocols