Isolation and culture of neonatal mouse calvarial osteoblasts.
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.
University of Rochester Medical Center, Rochester, NY, USA.
Jennifer H
JH
Jonason
University of Rochester Medical Center, Rochester, NY, USA.
Regis J
RJ
O'Keefe
D000818:Animals; D002469:Cell Separation; D006651:Histocytochemistry; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D061251:Primary Cell Culture; D012886:Skull / Q000166:cytology*; D014161:Transduction, Genetic