Fig. 4.

Depletion of pluripotent cells by K312 prevents teratoma formation by differentiated H9 hESCs. H9 hESCs were differentiated by retinoic acid (RA) treatment for 12 days. (A) Morphological differences between undifferentiated and RA-treated hESCs observed using a microscope. (B) Fluorescence-activated cell sorting of K312-high and -low cell populations from RA-treated H9 hESCs. (C) Immunoblotting analysis of the expression of the target of K312, Oct4, and Nanog in K312-high or K312-low cells. (D and E) Teratoma formation by K312-high and -low cells. Control mouse embryonic fibroblasts (MEFs), K312-low cells, or K312-high cells were injected into each side of the mouse testes. (D) Teratoma formation observed 8 weeks after cell transplantation. (E) Differences in the teratoma volume and weight between the groups. Error bars indicate ± SEM. **P < 0.01, Student’s t-test. (F) Representative hematoxylin and eosin (H&E) staining images of the testes sections. (G) RT-PCR analysis of the expression of the representative germ layer markers.