Figure 1.

Characterization of hypothalamus organoids

(A) Schematic overview of the hypothalamus differentiation protocol from hPSCs to NPCs and mature cells including DNs. hPSCs, human pluripotent stem cells; NPCs, neural progenitor cells; DNs, dopaminergic neurons.

(B) IF co-staining in D15 monolayer neurons for βIII Tubulin and RFP. Hoechst was used as a nuclear marker. Scale bar: 100 μm. IF, immunofluorescence; D15, day 15, RFP, red fluorescent protein.

(C) Fluorescent image of D20 organoids showing expression of the TH-TdTomato reporter. Scale bar: 250 μm. TH, tyrosine hydroxylase.

(D) IHC stains for TH and RFP for DNs, hypothalamic and ventral forebrain markers, OTP and NKX2-1, and hypothalamus-specific neuronal markers, POMC and TRH, in D80 hypothalamus organoids. TH + RFP co-staining was used for validation of the TH-TdTomato reporter in DNs, while TH + OTP co-staining showed a diversity of cell types. Hematoxylin was used as a nuclear marker in all images. Scale bars: 50 μm. IHC, immunohistochemistry.

See also Figure S1.