The development of genetic tools allowed for the validation of the pro-aging and pro-disease functions of senescent cells in vivo. These discoveries prompted the development of senotherapies—pharmaceutical interventions aimed at interfering with the detrimental effect of senescent cells—that are now entering the clinical stage. However, unequivocal identification and examination of cellular senescence remains highly difficult because of the lack of universal and specific markers. Here, to overcome the limitation of measuring individual markers, we describe a detailed two-phase algorithmic assessment to quantify various senescence-associated parameters in the same specimen. In the first phase, we combine the measurement of lysosomal and proliferative features with the expression of general senescence-associated genes to validate the presence of senescent cells. In the second phase we measure the levels of pro-inflammatory markers for specification of the type of senescence. The protocol can help graduate-level basic scientists to improve the characterization of senescence-associated phenotypes and the identification of specific senescent subtypes. Moreover, it can serve as an important tool for the clinical validation of the role of senescent cells and the effectiveness of anti-senescence therapies.

遗传工具的开发可以验证体内衰老细胞的促衰老和促疾病功能。这些发现促进了衰老疗法（旨在干扰衰老细胞有害影响的药物干预）的发展，目前已进入临床阶段。然而，由于缺乏通用和特异性标记物，明确识别和检查细胞衰老仍然非常困难。在这里，为了克服测量单个标记的限制，我们描述了详细的两阶段算法评估，以量化同一样本中的各种衰老相关参数。在第一阶段，我们将溶酶体和增殖特征的测量与一般衰老相关基因的表达结合起来，以验证衰老细胞的存在。在第二阶段，我们测量促炎标记物的水平，以指定衰老类型。该协议可以帮助研究生水平的基础科学家改善衰老相关表型的表征和特定衰老亚型的识别。此外，它可以作为临床验证衰老细胞的作用和抗衰老疗法有效性的重要工具。