FIG. 1.

Characterization of ESCs, ntESCs, and iPSCs. The purchased ES, ntES, and iPS cell lines were cultured for five or six generations before being used for experiments. The six pluripotent cell lines underwent 25–30 generations. Cell morphology resembled that of the ESCs. The expression of pluripotency markers was observed. (A) ESC-like cloned cells. (B) Represententive AP staining of iPSCs. (C) SSEA-1. (D) Eight-cell blastocyst injection of iPSCs. The iPSCs were delivered into embryos at the eight-cell stage and cultured in vitro for 48 h. The chimeric rate of the ICM blastocyst was 86.0±8.4% (n=10), where chimeric rate=ICM fluorescence area/ICM total area (the analysis was carried out with OLYMPUS cellSens Standard Vers1.5). (E) iPSCs were GFP positive, indicating successful expression of Oct4-GFP. (F) Karyotype analysis. The iPS2-19 and iPS2-33 cells at passage 30 showed a normal 40 XY karyotype.