Fig. 7: In vivo analysis of midbrain patterned cells transplanted into a Parkinson’s disease rat model.

a Overview of the in vivo study. Created with BioRender.com. b Amphetamine-induced rotational asymmetry. Two-way repeated measures ANOVA followed by Sidak’s multiple comparison test; time: P < 0.0001; treatment: P = 0.0009. **P = 0.0072 and ****P < 0.0001 vs. the same group at week −1. £P = 0.0225 vs. H9 group at the same time point. The data are presented as the mean ± SEM. 6-OHDA group n = 6, H9 group n = 9, and 4X group n = 9 rats. c Cylinder test. Two-way repeated measures ANOVA followed by Sidak’s multiple comparison test. Time x treatment: contra: P = 0.0039; ****P < 0.0001 vs. the same group at week −1. $$$P = 0.0003 vs. the 6-OHDA group at the same time point. ££P = 0.0048 vs. the H9 group at the same time point. The data are presented as the mean ± SEM. 6-OHDA group n = 4, H9 group n = 6, and 4X group n = 7 rats. d Representative photos of coronal sections H9 and 4X cell-transplanted groups for TH. Scale bars, 50 µm. e Estimated numbers of TH-positive cells in the grafts. f The yield of TH-positive neurons per 100,000 grafted cells. g TH-positive graft volume. h The estimated percentage of TH-positive cells within the HNA cells. The data in e-h are mean ± SEM. n = 9 rats per group. An unpaired two-tailed t test was used to compare groups in (e, f, h) and a Mann–Whitney two-tailed test for (g). i Representative immunofluorescence images of cells double-positive for TH/FOXA2, TH/LMX1A, TH/EN1 (n = 3 rats), TH/GIRK2 and TH/CALB1 (n = 9 rats) within H9 and 4X cell grafts. Scale bars, 50 μm. j Quantitative analysis of the immunofluorescence data showing the percentages of GIRK2/TH and CALB1/TH double-positive cells within TH cells in H9 and 4X cell grafts. The data are mean percentage ± SD (CALB1, H9: n = 8, 4X: n = 9; GIRK2, H9: n = 8, 4X: n = 7 rats).