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.

Original languageEnglish
Title of host publicationSkeletal Development and Repair
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Number of pages11
ISBN (Print)9781627039888
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Bone
  • Calvaria
  • Maturation
  • Mineralization
  • Mouse
  • Osteoblast


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