新生小鼠颅骨成骨细胞的分离与培养。
Isolation and culture of neonatal mouse calvarial osteoblasts.
作者信息
Jonason Jennifer H, O'Keefe Regis J
机构信息
University of Rochester Medical Center, Rochester, NY, USA.
出版信息
Methods Mol Biol. 2014;1130:295-305. doi: 10.1007/978-1-62703-989-5_22.
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.
摘要
本章描述了新生小鼠颅骨成骨细胞的分离和培养。这个原代细胞群体是通过对颅骨骨基质进行顺序酶消化获得的，并且能够在体外分化为成熟的成骨细胞，这些成骨细胞会沉积胶原蛋白细胞外基质并形成矿化骨结节。培养物的成熟可以通过基因表达分析以及碱性磷酸酶存在情况或基质矿化的染色来监测。因此，这个培养系统提供了一个强大的模型，用于测试各种实验条件，例如基因表达的操纵，如何影响成骨细胞的成熟和/或功能。
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