Figure 7.

ALS iPSC-Derived M2 Macrophages Sustain the Expression of Treg Functional Markers CD25 and Foxp3

Tregs purified from blood of ALS patients were either cultured alone or co-cultured with iPSC-derived M2 cells at a ratio of 1:1/2 (Tregs:M2). After 2 days, T-cells were collected for flow cytometric analyses.

(A) Representative plots showed that the percentage of CD4⁺CD25⁺Foxp3⁺ Tregs increased in a dose-dependent fashion after being co-cultured with M2 cells derived from a representative ALS C9 iPSC.

(B) Similar to CRT M2 cells (n = 3), M2 cells from ALS C9 iPSCs (n = 3) and sporadic ALS iPSCs (n = 4) increased the percentage of CD4⁺CD25⁺Foxp3⁺ Tregs. ∗∗∗p < 0.001 vs. ALS Tregs alone.

(C and D) M2-rescued CD4⁺CD25⁺ Tregs were isolated from M2 and ALS Tregs co-cultures. These M2-rescued Tregs (rTregs_(M2)) were then co-cultured with CFSE-stained responder Teffs from ALS patients for 5 days at a ratio of 1:1/16 (Tresp : M2-rescued Tregs). After co-cultured with Tregs rescued by ALS M2 cells, proliferation of ALS Tresp was lower than ALS Tresp alone (D) (^#p < 0.05, n = 3). Representative proliferation plots were shown in C.