Fig. 4: MPCs enhance the immunity and survival in the MB49 orthotopic bladder cancer model.

A Experimental workflow for selecting potent MPCs in the mouse bladder cancer model. B Experimental schematic: Mice were implanted with MB49 bladder tumors on day 0 and received low frequency intravesical MPC (MPC_LF) or DMEM control on days 2, 9, and 16, or MPC on days 2, 5, 8, 11, 14, and 17. On day 20, mice were euthanized, and tumor single cell suspensions were stained and analyzed by flow cytometry. C Representative images of mouse bladders on day 20. D Uniform Manifold Approximation and Projection (UMAP) analysis of tumor and immune cell components in DMEM-treated control and MPC17-treated mouse bladders. E Histopathological images of H&E-stained mice bladder tumors comparing Control and MPC17 at 20X magnification. The infiltration score increases from 2.52 ± 0.98 to 3.13 ± 0.78 (p = 0.0498, Student’s T test). Scale bar, 50 μm. Representative images of n = 8 for DMEM, n = 10 for MPC17, mouse biological replicates. Comparison of MPC-treated versus DMEM-treated bladders for: F WBC (CD45⁺) percent of total live cells, G representative flow cytometry plots of the immune cell population, H T cell (CD45⁺CD3⁺) as a percentage of total live cells, I CD4⁺ T cell (CD45⁺CD3⁺CD4⁺) as a percentage of total live cells, J CD8⁺ T cell (CD45⁺CD3⁺CD8⁺) as a percentage of total live cells, K B cell (CD45⁺CD19⁺) percent of total live cells, L NK cell (CD45⁺NK1.1⁺) as a percentage of total live cells, and M dendritic cell (CD45⁺CD11c⁺) as a percentage of total live cells. Data represent mean ± SE. P values from flow analysis were derived by the Kruskal-Wallis test followed by Dunn’s test, ns P > 0.05; *P < 0.05; **P < 0.01. Mouse biological replicates were used, with n = 8 for DMEM, MPC8, and MPC14; n = 10 for MPC15 and MPC17, all mice from two batches combined (B-M). N Experimental schematic: Mice were implanted with MB49 bladder tumors on day 0 and received intravesical MPC_LF or DMEM control on days 2, 9, 16, 23, and 30, or MPC on days 2, 5, 8, 11, 14, 17, 20, 23, 26, and 29. Survival of mice treated with (O) intravesical MPC and (P) MPC_LF. Statistical significance was assessed using the Kruskal-Wallis with multiple comparisons test. ns P > 0.05; ∙ P < 0.1; * P < 0.05; ** P < 0.01. Mice sample size: n = 15 for all survival studies, with the control group from the same experiment (N-P). Source data are provided as a Source Data file.