Fig. 1

In vitro generation of hiPSC-derived mDA precursors. (A) Human fibroblasts were obtained from a skin biopsy following an abdominoplasty performed on a healthy 28-year-old woman. (B) Dermal fibroblasts were reprogrammed into hiPSCs using mRNA transfection factors. (C) Schematic representation of the different stages (specification, expansion, and neuronal maturation) involved in the differentiation of hiPSCs into mesencephalic dopaminergic precursors. At the end of the differentiation stage on day 26 (D26), generated mDA precursors were generated and used for transplantation. (D) Representative images of immunolabeling on day 26, showing the expression of floor plate specification markers (OTX2, FOXA2, LMX1A), as well as DA precursor markers (EN2, NURR1 and PITX3), DA neuronal marker (TH) and the neural progenitor identity marker (NESTIN). Cell nuclei were counterstained with DAPI (blue). Scale bars: 50 μm. (E) Quantitative analysis of different markers expressed by DA precursors at day 26. Data are presented as mean ± SD (n = 3 independent experiments, with ≥ 8000 cells analyzed per experiment)