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Rapid detection of senescent mesenchymal stromal cells by a fluorescent probe.

Despite intense interest in human mesenchymal stromal cells (MSCs), monitoring its progressive occurrence of senescence has been hindered by the lack of efficient detection tools. Here, we report the discovery of a novel MSC senescence-specific fluorescent probe (CyBC9) identified by a high-throughput screen. Compared with the prototypical senescence-associated β-Gal (SA-β-gal) staining, the CyBC9 assay is rapid (2h) and non-toxic thus can be applied to live cells. We show that CyBC9 is able to stain early and late senescent populations both in monolayer and microcarrier-based cultures. Lastly, to investigate the mechanism of CyBC9, we performed co-localization assays and found that CyBC9 is accumulated in mitochondria of senescent MSCs presumably due to the loss of membrane potential. Taken together, we expect that CyBC9 will be a useful tool to ameliorate cell therapy through rapid and early screening of senescent phenotypes in clinically relevant MSCs. This article is protected by copyright. All rights reserved.