Abstract
This chapter describes the isolation and culture of neonatal mouse calvarial osteoblasts. This primary cell population is obtained by sequential enzymatic digestion of the calvarial bone matrix and is capable of differentiating in vitro into mature osteoblasts that deposit a collagen extracellular matrix and form mineralized bone nodules. Maturation of the cultures can be monitored by gene expression analyses and staining for the presence of alkaline phosphatase or matrix mineralization. This culture system, therefore, provides a powerful model to test how various experimental conditions, such as the manipulation of gene expression, may affect osteoblast maturation and/or function.
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Acknowledgements
The authors would like to thank Yongchun Zhang, Donna Hoak, and Tzong-jen Sheu for technical assistance. This work was supported by Public Health Service Grants RO1 AR053717, P50 AR054041, and P30 AR061307.
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Jonason, J.H., O’Keefe, R.J. (2014). Isolation and Culture of Neonatal Mouse Calvarial Osteoblasts. In: Hilton, M. (eds) Skeletal Development and Repair. Methods in Molecular Biology, vol 1130. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-989-5_22
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DOI: https://doi.org/10.1007/978-1-62703-989-5_22
Publisher Name: Humana Press, Totowa, NJ
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