Fig. 4.

Treatment of patient fibroblast-derived NPCs with AAV9.MECP2 restores differentiation potential into astrocytes that are supportive of neuron morphology and survival.

NPCs were transduced with AAV9.MECP2 and subsequently differentiated into astrocytes. (A) Cells were stained with the NPC marker, nestin, and an astrocyte marker, GFAP. (B) Quantification of GFAP and nestin staining intensity following differentiation. Transduction with AAV9.MeCP2 improved differentiation potential as indicated by reduced nestin and increased GFAP staining (scale bar: 100 ​μM). Astrocytes were seeded in co-culture with GFP+ ​neurons (black) and imaged 3 days later. (C) Representative images of neurons co-cultured with differentiated astrocytes. (D) Quantification of survival and neuronal morphology (scale bar: 200 ​μM). AAV9.MeCP2 significantly improved survival and neuronal morphology in the TCF4 deletion line (TCF4-3). Dotted line represents average control values. Data were generated from at least 3 independent experiments. Statistical analysis was performed using Student's T-test comparing the mean of untreated vs treated TCF4 cell lines. ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001, ∗∗∗∗p ​< ​0.0001.