Figure 1.

Purification and characterization of hiNSCs derived from iPSC

(A) Fluorescence-activated cell sorting (FACS) analysis of hiNSCs. CD184⁺/CD44⁻/CD271⁻sorting captured approximately 75.7% of the hiNSC population (top). After 4 months, approximately 89.44% of these previously sorted hiNSCs retained CD184 expression (bottom), indicating stability in the expression profile over time. (B) Immunocytochemistry staining for NSC markers Nestin, PAX6, SOX1, and SOX2 alongside CD184, a marker for migration capacity. Notably, the absence of OCT3/4, typically found in induced iPSCs, confirms the differentiated nature of the NSCs. Scale bar, 50 μm. (C) Cell migration assay. CD184⁺ hiNSCs grew and migrated primarily toward chemoattractant SDF-1α (100 ng/mL) (yellow arrow; left chamber) after 72-h incubation. Scale bar, 100 μm. (D) hiNSC transwell co-culture assay. Left graph: intracellular IDUA activity was significantly increased in MPS I-NSCs cross-corrected (light gray) with hiNSCs (dark gray), compared to uncorrected MPS I-NSCs (black). Right graph: GAG levels were significantly reduced in cross-corrected MPS I-NSCs, while GAG levels remained 2-fold higher in uncorrected MPS I-NSCs, with minimal amounts of enzyme activity as seen in the left graph. Data from biochemical assays are presented as mean ± SD. Statistical significance is denoted by asterisks (∗∗p < 0.01; ∗∗∗∗p < 0.0001; ns, not significant).