Kinetic Tracking of Therapy-Induced Senescence Using the Real-Time Cell Analyzer Single Plate System

Abstract

In recent years, terminal growth arrest, that is, senescence, especially therapy-induced senescence (TIS), has become a major subject in cancer research and several fields of life sciences. Senescence is characterized by a specific set of morphological and biochemical changes. However, methods that evidence senescence induction are still very limited and show large variation between individual examiners. Most notably, these assays are classical endpoint assays, and, therefore, screening for senescence is time consuming and expensive. Here, we describe an efficient, simple, and objective method to screen for TIS over time by modifying the Real-Time Cell Analyzer SP system, thus enabling to pin point the induction of senescence. This method continuously detects the cell's impedance in each well of a 96-microwell plate that allows to observe increment of cell size, a hallmark feature of cellular senescence. This technique is suitable for high-throughput TIS screening by measuring several compounds, small molecules, and/or cell lines simultaneously.