Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues
Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues
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DOI：
10.3791/53565
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发表时间：
2016-01-01
影响因子：
1.2
通讯作者：
Gasser, Stephan
中科院分区：
文献类型：
--
作者：
Khan, Muznah;Gasser, Stephan
原代细胞直接来源于组织，被认为比已建立的细胞系更能代表体内细胞的生理状态。然而，原代细胞培养物通常具有有限的寿命，并且需要频繁地重建。成纤维细胞是一种容易获得的原代细胞来源。在这里，我们讨论了一个简单而快速的实验程序，建立原代成纤维细胞培养从耳朵和尾巴的小鼠。该方案可用于建立在室温下储存长达10天的耳的原代成纤维细胞培养物。当仔细遵循该方案时，尽管在某些情况下使用了长时间储存的非无菌组织，但不太可能发生污染。成纤维细胞在培养中迅速增殖，并且在经历复制性衰老之前可以扩增到相当大的数量。
Primary cells are derived directly from tissue and are thought to be more representative of the physiological state of cells in vivo than established cell lines. However, primary cell cultures usually have a finite life span and need to be frequently re-established. Fibroblasts are an easily accessible source of primary cells. Here, we discuss a simple and quick experimental procedure to establish primary fibroblast cultures from ears and tails of mice. The protocol can be used to establish primary fibroblast cultures from ears stored at RT for up to 10 days. When the protocol is carefully followed, contaminations are unlikely to occur despite the use of non-sterile tissue stored for extended time in some cases. Fibroblasts proliferate rapidly in culture and can be expanded to substantial numbers before undergoing replicative senescence.