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. 2024 Aug 22;25(16):9115. doi: 10.3390/ijms25169115

Figure 1.

Figure 1

Schematic illustration outlining the timeline and conditions of the newly established protocol for the generation of hDPSC-derived SGN-like cells. The hDPSCs were exposed to SB/LDN for 4 days in vitro, followed by treatment with SB/BMP4 for an additional 3 days in vitro to generate neurospheres in ultra-low attachment (ULA) plates. The generated neurospheres were then plated on a Geltrex-coated surface and differentiated following differentiation paradigms 1 or 2. For paradigm 1, neurospheres were exposed to ATRA/SHH/CHIR until 13 DIV and then to BDNF/NT3 until 21 DIV. For paradigm 2, neurospheres were only exposed to BDNF/NT3 for the same period in vitro. The culture period was extended to 32 DIV in paradigm 2 to assess the effect of a prolonged in vitro maturation. Abbreviations: DIV: day in vitro; SB: SB431542: TGFb inhibitor; LDN: LDN-193189: bone morphogenetic protein (BMP) pathway inhibitor; BMP4: bone morphogenetic protein 4, ATRA: all trans retinoic acid; SHH: Sonic hedgehog; BDNF: brain-derived neurotrophic factor; NT3: neurotrophin 3.