Protocol

Skeletal Development and Repair

Volume 1130 of the series Methods in Molecular Biology pp 295-305

Date:

Isolation and Culture of Neonatal Mouse Calvarial Osteoblasts

  • Jennifer H. JonasonAffiliated withUniversity of Rochester Medical Center
  • , Regis J. O’KeefeAffiliated withUniversity of Rochester Medical Center

* Final gross prices may vary according to local VAT.

Get Access

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.

Keywords

Mouse Calvaria Osteoblast Bone Maturation Mineralization