Skip to main content
. 2023 Jul 12;17:1202027. doi: 10.3389/fnins.2023.1202027

Figure 1.

Figure 1

Characterization of mDA progenitors and neurons induced using the neurosphere-based differentiation protocol. (A) Overview of the differentiation protocol using chemically transitional embryoid-body-like state (CTraS) method from human iPSCs to dopaminergic neurons. The treatment with three chemicals of iPSCs induces CTraS. (B) qRT-PCR analysis of anteroposterior markers in neurospheres derived from iPSCs (All samples n = 3, mean ± standard error of mean [SEM]). Values were normalized to ACTB and were analyzed using the comparative (ΔΔCt) method. *p < 0.05, **p < 0.01. (C–F) Immunostaining analysis of mDA progenitors on day 20. Immunofluorescence images of mDA progenitor markers (FOXA2, LMX1A, NURR1, CORIN, ALCAM, and LRTM1) (C,E) and their positivity rates (D,F). (G,H) Immunostaining analysis of the differentiated mDA neurons in vitro on day 36. Immunofluorescence images of the mDA neuron markers (G) and the positive rate of GIRK2, an A9 mDA neuron marker, and Calbindin, an A10 mDA neuron marker for TH-positive cells (H). Bars, 50 μm. mDA, midbrain dopaminergic; qRT-PCR, quantitative real-time Polymerase Chain Reaction; iPSCs, induced pluripotent stem cells; N.D., not detected.