Equivalency of Nuclear Transfer‐Derived Embryonic Stem Cells to Those Derived from Fertilized Mouse Blastocysts

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Abstract
Therapeutic cloning, whereby nuclear transfer (NT) is used to generate embryonic stem cells (ESCs) from blastocysts, has been demonstrated successfully in mice and cattle. However, if NT‐ESCs have abnormalities, such as those associated with the offspring produced by reproductive cloning, their scientific and medical utilities might prove limited. To evaluate the characteristics of NT‐ESCs, we established more than 150 NT‐ESC lines from adult somatic cells of several mouse strains. Here, we show that these NT‐ESCs were able to differentiate into all functional embryonic tissues in vivo. Moreover, they were identical to blastocyst‐derived ESCs in terms of their expression of pluripotency markers in the presence of tissue‐dependent differentially DNA methylated regions, in DNA microarray profiles, and in high‐coverage gene expression profiling. Importantly, the NT procedure did not cause irreversible damage to the nuclei. These similarities of NT‐ESCs and ESCs indicate that murine therapeutic cloning by somatic cell NT can provide a reliable model for preclinical stem cell research.