A deep single cell mass cytometry approach to capture canonical and noncanonical cell cycle states

Meelad Amouzgar, Patricia Favaro, Daniel Ho, Trevor Bruce and Sean C. Bendall ()
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Meelad Amouzgar: Stanford University
Patricia Favaro: Stanford University
Daniel Ho: Stanford University
Trevor Bruce: Stanford University
Sean C. Bendall: Stanford University

Nature Communications, 2025, vol. 16, issue 1, 1-14

Abstract: Abstract The cell cycle (CC) underpins diverse cell processes like cell differentiation, cell expansion, and tumorigenesis but current single-cell (sc) strategies study CC as: coarse phases, rely on transcriptomic signatures, use imaging modalities limited to adherent cells, or lack high-throughput multiplexing. To solve this, we develop an expanded, Mass Cytometry (MC) approach with 48 CC-related molecules that deeply phenotypes the diversity of scCC states. Using Cytometry by Time of Flight, we quantify scCC states across suspension and adherent cell lines, and stimulated primary human T cells. Our approach captures the diversity of scCC states, including atypical CC states beyond canonical definitions. Pharmacologically-induced CC arrest reveals that perturbations exacerbate noncanonical states and induce previously unobserved states. Notably, primary cells escaping CC inhibition demonstrated aberrant CC states compared to untreated cells. Our approach enables deeper phenotyping of CC biology that generalizes to diverse cell systems with simultaneous multiplexing and integration with MC platforms.

Date: 2025
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DOI: 10.1038/s41467-025-63883-4

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