Figure 7.
Increased Dopaminergic Differentiation of NSCs Independent on Prior Microglial Activation and Identification of TNFα, IL-1β, and IGF1 as Potential Mediators
(A–C) Immunocytochemical staining and quantification of hVM1-Bcl-XL NSC-derived neurons after co-culture differentiation with pro- and anti-inflammatory activated BV2 microglia for (A) TH+ neurons/total cell count, (B) TH+ neurons/β-tubulin III+ neurons, and (C) β-tubulin III+ neurons/total cell count. One-way ANOVA, Dunnett's multiple comparison test with reference to control. Control, n = 20, N = 7; untreated BV2, n = 15, N = 5; LPS-treated BV2, n = 10, N = 4; IL-4 treated BV2, n = 12, N = 5.
(D) IGF1 ELISA; all groups, n = 4, N = 2, and (E) cytokine profiling; control, n = 6, N = 3; untreated BV2, LPS-treated BV2, and IL-4 treated BV2, n = 4, N = 2, of co-culture medium collected at day 3 of differentiation. One-way ANOVA, Tukey's multiple comparison test.
(F–H) Effect on TH+ neurons/total cell count of direct addition of recombinant human (F) TNFα, (G) IL-1β, and (H) IGF1 to hVM1-Bcl-XL NSCs during differentiation. One-way ANOVA, Dunnett's multiple comparison test with reference to control. All groups, n = 6, N = 2.
(I) Most efficient concentrations of recombinant TNFα, IL-1β, and IGF1 tested on iPSC-NSCs. One-way ANOVA, Dunnett's multiple comparison test with reference to control. All groups, n = 6, N = 2. Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, NS = not significant.
See Figures S6 and S7 for additional data.