Figure 3.

The partial conversion to the naive state enabled hiPSCs to differentiate into AVP neurons and hypothalamic organoid. (A) Scheme of the culture protocol. SFEBq using 201B7 cells recapitulated embryogenesis; on day 30, dorsal hypothalamic progenitor cells [PAX6+ (green), NKX2.1− (red)] (B), and on day 60, AVP precursor cells [OTP+ (green), BRN2+ (red)] (C), were sequentially identified. FOXG1 (white). (D) On day 150, differentiation into AVP neurons was achieved, with co-expression of pro-AVP components: AVP (green), NPII (red), and copeptin (white). Day 150 aggregates also expressed other hypothalamic markers such as CRF (E), NPY (F), AgRP (G), TRH (H), and Orexin (I). It is demonstrated that these cells are not only AVP neurons but also hypothalamic organoids. For all relevant panels, nuclear counterstaining was performed with DAPI (blue). Scale bars, (B) 100 µm, (C–I) 50 µm.