• Long-term culture of tonsil mesenchymal stem cells (TMSCs) leads to replicative senescence characterized by reduced proliferation and increased SA-β-gal activity.
• Senescent TMSCs exhibit upregulation of senescence markers p16, p53, and p21.
• RNA-seq and KEGG analysis reveal activation of the PI3K-Akt signaling pathway in senescent TMSCs.
• Increased p-Akt/Akt ratio in senescent TMSCs suggests PI3K-Akt pathway as a potential therapeutic target to delay senescence.