Monitoring senescent cell dynamics in wound healing using a p16-tdtomato mouse model.

Abstract

Although accumulating evidence implicates cellular senescence in acute wound healing, the precise roles of senescent cells within distinct cell lineages during this process remain elusive. To address this, we employed the p16-tdTomato reporter mouse model for labeling and isolating senescent cells from wound tissue. Longitudinal in vivo imaging monitoring revealed the temporal dynamics of tdTomato (tdTom) fluorescence, with signal detection as early as postoperative day 3, peaking by day 6. Utilizing an optimized tissue digestion protocol, we achieved high-viability FACS isolation of p16-expressing cells (tdTom+), which exhibited characteristic senescent cell morphology and marker expression. Single-cell analysis demonstrated that tdTom+ wound cells enriched with p16 expression were primarily characterized as fibroblasts and displayed common features of senescence. These findings validate the p16-tdTomato reporter mouse as a reliable model for identifying and isolating senescent cells from the wound microenvironment at single-cell resolution.