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. 2017 Oct 23;114(45):11974–11979. doi: 10.1073/pnas.1708710114

Fig. 1.

Fig. 1.

Generation of germ line-competent rat ESCs. (A) The compact colony morphology of rat ESCs at passage 16. (Scale bar, 100 μm.) (B) Alkaline phosphatase staining of rat ESCs at passage 16. (Scale bar, 100 μm.) (C) RT-PCR analysis of pluripotent markers expressed in two rat ESC lines (DAESC-1 and DAESC-2). Rat tail-tip fibroblast cells and H2O served as negative controls. (D) Immunostaining of OCT4, NANOG, SOX2, and SSEA-1 in rat ESCs. (Scale bar, 100 μm.) (E) Whole-genome sequencing analysis showed the genome integrity of DAESC-1 at passage 13. Relative copy number is plotted using log2 scale. Each point represents the average coverage of 500 kb. (F) Morphology of EBs formed from rat ESCs. (Scale bar, 100 μm.) (G) Teratoma formation of rat ESCs. Shown are histological sections of teratoma including adipose tissue for endoderm (black arrows indicated, Left), muscle for mesoderm (black arrow indicated, Middle), and neural epithelium rosettes for ectoderm (black arrows indicated, Right). (Scale bar, 500 μm.) (H) Coat chimera generated from rat ESCs by blastocyst injection. Black indicates the contribution of donor rat ESCs, and white indicates the host diploid F344 embryos. (I) Germ-line offsprings produced by the chimeric female rat mated with a wild-type SD male rat. Five germ-line offsprings were obtained (black pups as indicated by the red stars) among 13 F1 pups.