Long-term culture of mature midbrain organoids
(A) qRT-PCR showing mRNA expression of mature midbrain DN markers NURR1 and GIRK2 in D30 midbrain organoids as a fold change over hPSCs (n=3). ACTB was used as a housekeeping gene for normalization. Error bars represent SD.
(B) FACS analysis of dissociated D30 WA01 TH midbrain organoids showing a ~30% TH-TdTomato+ cell population.
(C) Tiled images showing TH-TdTomato reporter expression in a D35 WA01 TH midbrain organoid splatted on D16. Scale bar: 1 mm.
(D) IHC staining of a D80 BJ-SiPS TH midbrain organoid section indicating the presence of astrocytes via GFAP (brown) expression. The nuclei are counterstained with hematoxylin (light blue). Scale bar: 100 μm.
(E) IHC staining of a D200 BJ-SiPS TH midbrain organoid section showing TH (brown) and melanin (dark blue) expression. Arrows point to examples of regions in which melanin is localized to DNs. The nuclei are counterstained with hematoxylin (light blue). Scale bar: 100 μm.