Althubiti Mohammad, Macip Salvador
Mechanisms of Cancer and Aging Laboratory, Department of Molecular and Cell Biology, University of Leicester, University Road, LE1 7RH, Leicester, UK.
Cancer Research UK Leicester Centre, Leicester, UK.
Methods Mol Biol. 2017;1534:147-153. doi: 10.1007/978-1-4939-6670-7_14.
Senescence is a cellular process that is thought to have prognostic and therapeutic relevance in conditions such as cancer, aging, and fibrosis. However, current protocols for identifying senescent cells in vitro and in vivo have several drawbacks. Most markers used lack sufficient specificity and false positives and negatives in common. In addition, classical staining techniques often require lengthy protocols and do not offer objective quantification. Recently, several novel markers of senescence associated with the plasma membrane have been identified. Here, we propose to take advantage of these markers to define a customizable FACS-based protocol to detect senescent cells using antibodies tagged with fluorescence dyes. This method has the advantage of being fast and allowing quantitation. Furthermore, its specificity is increased using several markers simultaneously.
衰老 是一种细胞过程，被认为在癌症、衰老和纤维化等病症中具有预后和治疗相关性。然而，目前在体外和体内鉴定衰老细胞的方案存在若干缺陷。常用的大多数标志物缺乏足够的特异性，假阳性和假阴性情况常见。此外，经典染色技术通常需要冗长的流程，且无法提供客观的定量分析。最近，已鉴定出几种与质膜相关的新型衰老标志物。在此，我们建议利用这些标志物来定义一种基于流式细胞术的可定制方案，使用荧光染料标记的抗体来检测衰老细胞。该方法具有快速且可定量的优点。此外，同时使用多种标志物可提高其特异性。
