3D culture is more effective for iDAN direct reprogramming. (A) Schematic illustration describing the timeline of iDAN direct reprogramming of 3D culture. (B) The astrocytes and microglia seeded in AggreWellTM 800 plates sink over time and form neurospheres. (C) After 4 weeks of reprogramming, the expression of the dopaminergic neuron markers FOXA2 (red) and TH (green) is observed. (D) The colonies without microglia (−MG) and those with microglia (+MG) were cultured on the shaker for 7 and 14 days. (E) The expression of dopaminergic neuron markers in the (−MG) and (+MG) colonies is observed. FOXA2 (red), TH (green). Scale bar, 20 μm.
