A single-cell micro-trench platform for automatic monitoring of cell division and apoptosis after chemotherapeutic drug administration

Abstract

Cells vary in their dynamic response to external stimuli, due to stochastic fluctuations and non-uniform progression through the cell cycle. Hence, single-cell studies are required to reveal the range of heterogeneity in their responses to defined perturbations, which provides detailed insight into signaling processes. Here, we present a time-lapse study using arrays of micro-trenches to monitor the timing of cell division and apoptosis in non-adherent cells at the single-cell level. By employing automated cell tracking and division detection, we precisely determine cell cycle duration and sister-cell correlations for hundreds of individual cells in parallel. As a model application we study the response of leukemia cells to the chemostatic drug vincristine as a function of cell cycle phase. The time-to-death after drug addition is found to depend both on drug concentration and cell cycle phase. The resulting timing and dose-response distributions were reproduced in control experiments using synchronized cell populations. Interestingly, in non-synchronized cells, the time-to-death intervals for sister cells appear to be correlated. Our study demonstrates the practical benefits of micro-trench arrays as a platform for high-throughput, single-cell time-lapse studies on cell cycle dependence, correlations and cell fate decisions in general.

The effects of proliferation status and cell cycle phase on the responses of single cells to chemotherapy

“…Traditional assays of cellular response to chemotherapy involve analyzing the expression of proliferation markers in fixed cells, and therefore cannot link the proliferation outcomes to the prior states of the cells. Live microscopy that follows the response of individual cells to treatment over time has identified preexisting cell-state features (e.g., expression of MYC or cell cycle phase) that impact cellular outcomes in mammalian systems (Paek et al, 2016;Ryl et al, 2017;Chatzopoulou et al, 2018). Studies tracking multiple cell generations and sister cells over time as they respond to DNA damage and drug compounds suggest that multigeneration longterm memory features dominate cellular outcome choices (Arora et al, 2017;Chakrabarti et al, 2018;Korsnes and Korsnes, 2018;Wolff et al, 2018).…”