Passos João F, von Zglinicki Thomas
Henry Wellcome Laboratory for Biogerontology Research Newcastle University, Newcastle, UK.
Methods Mol Biol. 2007;371:33-44. doi: 10.1007/978-1-59745-361-5_4.
Cellular senescence, the ultimate and irreversible loss of replicative capacity of cells in primary culture, has been a popular model for studying the aging process. However, the replicative life span of human fibroblasts is heterogeneous even in clonal populations, with the fraction of senescent cells increasing at each population doubling, rather than all cells entering senescence simultaneously. Thus, the study of individual cells in a mass culture is of extreme importance to the understanding of replicative senescence. Cell sorting is a method that allows physical separation of cells with different characteristics when measured by flow cytometry. Here, we describe various methods by which cells that reach senescence early can be physically sorted out of a bulk of growing cells, and discuss how different methods can affect the posterior analysis of the sorted populations.
细胞衰老，即原代培养细胞复制能力的最终且不可逆丧失，一直是研究衰老过程的常用模型。然而，即使在克隆群体中，人类成纤维细胞的复制寿命也是异质的，衰老细胞的比例在每次群体倍增时都会增加，而不是所有细胞同时进入衰老状态。因此，对大规模培养中的单个细胞进行研究对于理解复制性衰老极为重要。细胞分选是一种通过流式细胞术测量时能够对具有不同特征的细胞进行物理分离的方法。在这里，我们描述了各种方法，通过这些方法可以从大量生长的细胞中物理分选早期达到衰老的细胞，并讨论不同方法如何影响分选群体的后续分析。
