Figure 3.

Consistent Positive Effect on Dopaminergic Differentiation Across Different Microglia Cell Lines

(A) Immunocytochemical staining of murine BV2 and human CHME microglia cells for the microglial marker Iba1. Scale bar: 50 μm.

(B–E) Immunocytochemical staining and quantification of hVM1-Bcl-X_L NSC-derived neurons after CHME and BV2 microglia co-culture differentiation for (B) TH⁺ neurons/total cell count, (C) TH⁺ neurons/β-tubulin III⁺ neurons, (D) total cell count, and (E) β-tubulin III⁺ neurons/total cell count. Control, n = 21, N = 8; BV2, n = 15, N = 5; CHME, n = 12, N = 5.

(F–I) Immunofluorescence staining and quantification of (F and I) GABA⁺ neurons/total cell count, (G) GABA⁺ neurons/β-tubulin III⁺ neurons, and (H and I) GFAP⁺-astrocytes/total cell count. Scale bar: 100 μm. Control, n = 6, N = 2; BV2, n = 6, N = 2; CHME, n = 6, N = 2.

(J) Immunofluorescence staining of neonatal and adult primary microglia (p. microglia) for the microglial marker Iba1. Scale bar: 100 μm.

(K–M) Quantification of (K) TH⁺ neurons/total cell count; control, n = 18, n = 4; neonatal p. microglia, n = 9, N = 2; adult p. microglia, n = 12, N = 4, (L) β-tubulin III⁺ neurons/total cell count, and (M) TH⁺ neurons/β-tubulin III⁺ neurons; control, n = 14, n = 4; neonatal p. microglia, n = 9, N = 2; adult p. microglia, n = 8, N = 2, after differentiation of hVM1-Bcl-X_L NSCs in co-culture with murine neonatal or adult p. microglia.

(N–P) Quantification of (N) TH⁺ neurons/total cell count, (O) β-tubulin III⁺ neurons/total cell count, and (P) TH⁺ neurons/β-tubulin III⁺ neurons after differentiating hVM1-Bcl-X_L NSCs co-cultured with murine CD11c⁺ or CD11c⁻ neonatal p. microglia. Control, n = 17, N = 6; CD11c⁺neo. p. microglia, n = 10, N = 4, CD11c⁻neo. p. microglia, n = 10, N = 4. Mean ± SEM. One-way ANOVA, Dunnett's multiple comparison test with reference to control. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

See Figures S2–S4 for additional data.