Figure 3

Passage solution with Ca²⁺ and without Mg²⁺ supports long-term culture and pluripotency of hPSCs.

The hiPSC 201B7 were cultured for 15 passages under the ESF-Fb-EzF condition. (a): Phsecontrast micrograph. (b–f): Immunocytochemistry showed that the cells expressed self-renewal markers, NANOG ((b): red), OCT3/4 ((c): red), SSEA4 ((c): red) and TRA 1–60 ((e): red), but not an early differentiation marker, SSEA1 ((f): red). The nuclei were stained with DAPI (blue). (g–h): Histological analysis with HE staining demonstrated that hiPSC-derived teratoma contained derivatives of all three germ layers: neural tube ((g): ectoderm), epitherial ((h): endoderm) and cartridge ((i): mesoderm). (j): FISH karyotype analysis showed a normal karyotype (46XX). Scale bars are 100 μm (a, g–i) or 200 μm (b–f).