Figure 3

Modified TERT mRNA delivery to T cells increased proliferative capacity and transient reduction of senescence-associated markers. (a) The clones of CD19 CAR T cells 7 days post transfection with TERT mRNA. (b) Growth curves of CD19 CAR T cells transfected with TERT mRNA, CI-TERT mRNA, three times in succession at 24 h intervals starting at PD 13, and the growth curves were repeated twice with each population cultured in triplicate compared with untreated CAR T cells. *P<0.05, ***P<0.001. (c) The cell cycle analysis was performed using flow cytometry (TERT mRNA group, CI-TERT mRNA group, GFP T group or CAR T group). (d) Only the percentage of total S-phase of each T cell line was statistically summarized. The percentage of total S-phase represents the proliferation of T cells. The data are presented as the mean±s.d. of results from three independent experiments. *P<0.05, ***P<0.001. (e, f) Quantification of senescent marker expressing T cells after modified TERT mRNA transfection three times in succession at 24 h intervals. The β-gal-positive T cells are shown in grayish-green in the microscope images. Before β-gal staining, the T cells were collected using centrifugation, washed with PBS and resuspended. The data are presented as the mean±s.d. of results from three independent experiments. *P<0.05, ***P<0.001