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. 2023 Jul 12;17:1202027. doi: 10.3389/fnins.2023.1202027

Figure 2.

Figure 2

Grafted mDA progenitors using a neurosphere-based protocol differentiate into mDA neurons in the brains of 6-OHDA-induced PD-model mice. (A) Representative immunofluorescence images of the CTraS-mediated grafts (upper panels) and sham-operated sample (lower panels) at 16 weeks after transplantation. Bars, 3 mm. R indicates the right side of the section. (B) Representative immunofluorescence images of grafted cells against SC-121, a human cytoplasmic marker, at 16 weeks after transplantation. Bars, 200 μm. (C,D) Immunostaining analysis of the graft at 16 weeks for mDA neuron-associated markers. Immunofluorescence images of indicated markers (C) and quantification of the marker-positive cells in HNA-positive cells (D). Bars, 50 μm. (E,F) Immunofluorescence images of mDA neuron markers (E), and quantification of marker-positive cells in HNA- and TH- double-positive cells (F). Bars, 100 μm. (G) Immunofluorescence images of subtype-specific mDA neuron markers. Bars, 10 μm. (H) Immunofluorescence images of astrocyte markers. Bars, 100 μm. (I) Immunofluorescence images of VLMC markers. The COL1A1 antibody recognizes both human and rodent COL1A1, and hCOL1A1 antibody recognizes human-specific COL1A1. Arrows indicate human iPSC-derived COL1A1-positive cells, and arrowheads indicate mouse-derived COL1A1-positive cells. Bars, 100 μm. mDA, midbrain dopaminergic; 6-OHDA, 6-hydroxydopamine; PD, Parkinson’s disease; CTraS chemically transitional embryoid-body-like state; HNA, human nuclear antigen; TH, Tyrosine hydroxylase; VLMC, vascular leptomeningeal-like cell.