We investigated whether neuronal cells differentiated from patient-derived hiPSC show abnormal cell phenotypes. First, we induced neural differentiation, by using non-adherent floating culture and then plated the cells on a poly-l-ornithine- and fibronectin-coated surface. We then compared the lengths of outgrown neurites and distance of cellular migration between control and patient-derived neurospheres. The neurites were visualized by immunocytochemical staining using the neuronal marker βIII-tubulin (Figure 2a). The neurite lengths were significantly shortened in patients' neurospheres (112±36 μm), compared with controls (204±19 μm; P=0.0345; Figure 2b). The nuclei were visualized by immunocytochemical staining using the neuronal marker, NeuN (Figure 2c). The migration distances were significantly decreased in neurospheres from patients (90±8 μm), compared with controls (133±7 μm; P=0.0032; Figure 2d). Therefore, the shortened neurite lengths in patients' samples cannot solely be explained by impaired cell migration.
