Isolation and Purification of Satellite Cells from Young Rats by Percoll Density Gradient Centrifugation
Isolation and Purification of Satellite Cells from Young Rats by Percoll Density Gradient Centrifugation
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DOI：
10.1007/978-1-4939-8897-6_6
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发表时间：
2019-01-01
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影响因子：
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通讯作者：
Mizunoya, Wataru
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文献类型：
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作者：
Matsuyoshi, Yuji;Akahoshi, Mariko;Mizunoya, Wataru
卫星细胞（SCs）是一种肌源性干细胞，在骨骼肌再生和肥厚中起重要作用。SCs原代培养可用于分析细胞功能；然而，用常规方法很难从幼鼠身上获得高纯度的sc。本研究的目的是建立一种分离幼年大鼠SC的纯化方法，并利用Percoll这一常用的细胞纯化工具对纯化过程进行定量评价。我们利用实时RT-qPCR和免疫细胞化学对Percoll密度梯度离心收集的SCs进行纯度鉴定。Percoll处理将从年轻大鼠中分离的SCs纯度提高到近90%，这与使用中年大鼠的常规方法所获得的纯度相当。
Satellite cells (SCs) are myogenic stem cells that play an important role in skeletal muscle regeneration and hypertrophy. Primary cultures of SCs are useful to analyze cell functions; however, it is difficult to obtain highly pure SCs from young rats with the conventional procedures. The purpose of this study is to establish a purification method for SC isolation from young rats and quantitatively evaluate the purification procedure employing Percoll, a common research tool to purify cells. We elucidated the purity of SCs collected by Percoll density gradient centrifugation using real-time RT-qPCR and immunocytochemistry for desmin. Percoll treatment increased the purity of SCs isolated from young rats to nearly 90%, which was comparable to that achieved with the conventional method using middle-aged rats.