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Rapid Detection of Senescent Mesenchymal Stromal Cells by a Fluorescent Probe
Author(s) -
Ang Joshur,
Lee YongAn,
Raghothaman Deepak,
Jayaraman Premkumar,
Teo Kim L.,
Khan Fahima J.,
Reuveny Shaul,
Chang YoungTae,
Kang NamYoung,
Oh Steve
Publication year - 2019
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201800691
Subject(s) - mesenchymal stem cell , fluorescence , stromal cell , chemistry , microbiology and biotechnology , biophysics , nanotechnology , biology , materials science , cancer research , physics , optics
Despite intense interest in human mesenchymal stromal cells (MSCs), monitoring of the progressive occurrence of senescence has been hindered by the lack of efficient detection tools. Here, the discovery of a novel MSC senescence‐specific fluorescent probe (CyBC9) identified by a high‐throughput screen is reported. Compared with the prototypical senescence‐associated β‐galactosidase (SA‐β‐gal) staining, the CyBC9 assay is rapid (2 h) and nontoxic and can thus be applied to live cells. It is shown that CyBC9 is able to stain early and late senescent populations both in monolayer‐ and in microcarrier‐based cultures. Finally, to investigate the mechanism of CyBC9, colocalization assays are performed and it is found that CyBC9 is accumulated in the mitochondria of senescent MSCs presumably due to the loss of membrane potential. Taken together, it is expected that CyBC9 will be a useful tool to ameliorate cell therapy through rapid and early screening of senescent phenotypes in clinically relevant MSCs.