Gal Hilah, Porat Ziv, Krizhanovsky Valery
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Flow Cytometry Unit, Life Sciences Core Facilities, Weizmann Institute of Science, Rehovot, Israel.
Methods Mol Biol. 2019;1896:107-117. doi: 10.1007/978-1-4939-8931-7_11.
Cellular senescence is a permanent growth arrest mechanism triggered by various forms of stress. Senescent cells accumulate in the mammalian organism with age and are present at sites of tissue damage and age related pathologies. However, the characterization of senescence cells in vivo is currently limited and the need for new technologies to detect and monitor the senescence state in vivo has greatly increased. Here we demonstrate the use of the ImageStreamX as a powerful method for detection and quantification of senescent cells at distinct tissues and cell subpopulations. The identification of senescent cells using ImageStreamX enables the use of a combination of several senescence-related markers, together with the commonly used senescence-associated beta-galactosidase assay. These can be combined with the use of other molecular features typical of senescence cells, such as the γH2AX foci, indicating the activation of DNA damage response. This novel method offers a feasible solution to quantify senescent cells in vivo, in a comprehensive manner. Such quantification is necessary in order to understand the role of cellular senescence in aging and disease.
细胞衰老一种由各种形式的应激触发的永久性生长停滞机制。衰老细胞会随着年龄增长在哺乳动物体内积累,并存在于组织损伤部位和与年龄相关的病理部位。然而,目前对体内衰老细胞的表征有限,因此对用于检测和监测体内衰老状态的新技术的需求大幅增加。在此,我们展示了使用ImageStreamX作为一种强大的方法,用于检测和定量不同组织和细胞亚群中的衰老细胞。使用ImageStreamX鉴定衰老细胞能够结合使用几种与衰老相关的标志物,以及常用的衰老相关β-半乳糖苷酶检测方法。这些可以与衰老细胞典型的其他分子特征(如γH2AX焦点,表明DNA损伤反应的激活)结合使用。这种新方法为全面定量体内衰老细胞提供了一种可行的解决方案。为了了解细胞衰老在衰老和疾病中的作用,这种定量是必要的。