Biran Anat, Zada Lior, Abou Karam Paula, Vadai Ezra, Roitman Lior, Ovadya Yossi, Porat Ziv, Krizhanovsky Valery
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, 76100, Israel.
Flow Cytometry Unit, Biological Services Department, Weizmann Institute of Science, 76100, Rehovot, Israel.
Aging Cell. 2017 Aug;16(4):661-671. doi: 10.1111/acel.12592. Epub 2017 Apr 28.
Senescent cells are present in premalignant lesions and sites of tissue damage and accumulate in tissues with age. In vivo identification, quantification and characterization of senescent cells are challenging tasks that limit our understanding of the role of senescent cells in diseases and aging. Here, we present a new way to precisely quantify and identify senescent cells in tissues on a single-cell basis. The method combines a senescence-associated beta-galactosidase assay with staining of molecular markers for cellular senescence and of cellular identity. By utilizing technology that combines flow cytometry with high-content image analysis, we were able to quantify senescent cells in tumors, fibrotic tissues, and tissues of aged mice. Our approach also yielded the finding that senescent cells in tissues of aged mice are larger than nonsenescent cells. Thus, this method provides a basis for quantitative assessment of senescent cells and it offers proof of principle for combination of different markers of senescence. It paves the way for screening of senescent cells for identification of new senescence biomarkers, genes that bypass senescence or senolytic compounds that eliminate senescent cells, thus enabling a deeper understanding of the senescent state in vivo.
衰老细胞存在于癌前病变和组织损伤部位，并随着年龄的增长在组织中积累。衰老细胞的体内鉴定、定量和表征是具有挑战性的任务，限制了我们对衰老细胞在疾病和衰老中作用的理解。在这里，我们提出了一种在单细胞水平上精确量化和鉴定组织中衰老细胞的新方法。该方法将衰老相关β-半乳糖苷酶检测与细胞衰老分子标记和细胞身份标记的染色相结合。通过利用将流式细胞术与高内涵图像分析相结合的技术，我们能够对肿瘤、纤维化组织和老年小鼠组织中的衰老细胞进行定量。我们的方法还发现老年小鼠组织中的衰老细胞比非衰老细胞大。因此，该方法为衰老细胞的定量评估提供了基础，并为不同衰老标记物的组合提供了原理证明。它为筛选衰老细胞以鉴定新的衰老生物标志物、绕过衰老的基因或消除衰老细胞的衰老溶解化合物铺平了道路，从而能够更深入地了解体内的衰老状态。
