Hewitt Graeme, von Zglinicki Thomas, Passos João F
Ageing Research Laboratories, Centre for Integrated Systems Biology of Ageing and Nutrition, Institute for Ageing and Health, Newcastle University, Newcastle, UK.
Methods Mol Biol. 2013;1048:31-47. doi: 10.1007/978-1-62703-556-9_4.
Cellular senescence is the irreversible loss of proliferative potential and is accompanied by a number of phenotypic changes. First described by Hayflick and Moorhead in 1961, it has since become a popular model to study cellular aging. The replicative lifespan of human fibroblasts is heterogeneous even in clonal populations, with the fraction of senescent cells increasing with each population doubling (PD). Thus, the study of individual cells in mass culture is necessary in order to properly understand senescence and its associated phenotype. Cell sorting is a process that allows the physical separation of cells based on different characteristics which can be measured by flow cytometry. Here, we describe various methods by which senescent cells can be sorted from mixed cultures and discuss how different methods impact on the posterior analysis of sorted populations.
细胞衰老指细胞增殖潜能的不可逆丧失，并伴有一系列表型变化。1961年由海弗利克和穆尔黑德首次描述，此后它成为研究细胞衰老的常用模型。即使在克隆群体中，人类成纤维细胞的复制寿命也是异质性的，衰老细胞的比例随着群体倍增数（PD）的增加而增加。因此，为了正确理解衰老及其相关表型，有必要对大规模培养中的单个细胞进行研究。细胞分选是一个基于不同特征对细胞进行物理分离的过程，这些特征可通过流式细胞术进行测量。在此，我们描述了从混合培养物中分选衰老细胞的各种方法，并讨论了不同方法如何影响分选群体的后续分析。
