ORIGINAL ARTICLE

Cell Research (2007): 80-87
© 2007 IBCB, SIBS, CAS All rights reserved 1001-0602/06 $ 30.00
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Establishment of customized mouse stem cell lines by sequential nuclear transfer

Chunli Zhao^{1, 2, *}, Ruqiang Yao^{1, 4,*}, Jie Hao^{1, 4}, Chenhui Ding^{1, 3}, Yong Fan², Xiangpeng Dai^{1, 4}, Wei Li^{1, 4}, Tang Hai^{1, 4}, Zichuan Liu^{1, 2}, Yang Yu^{1, 2}, Yingying Wang^{1, 3, 4}, Xiaojun Hou^{1, 4}, Weizhi Ji^{2, 3}, Qi Zhou^{1, 2}, Alice Jouneau^{1, 2, 5}, Fanyi Zeng^{2, 6}, Liu Wang^{1, 2}

*These two authors contribute equally to the work. Correspondence: Liu Wang1 and Fanyi Zeng2 1, 2Tel/ Fax: 86-10-62650042 1E-mail: qzhou@ioz.ac.cn; 2E-mail: zengfy@shsmu.edu.cn

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Therapeutic cloning, whereby embryonic stem cells (ESCs) are derived from nuclear transfer (NT) embryos, may play a major role in the new era of regenerative medicine. In this study we established forty nuclear transfer-ESC (NTESC) lines that were derived from NT embryos of different donor cell types or passages. We found that NT-ESCs were capable of forming embryoid bodies. In addition, NT-ESCs expressed pluripotency stem cell markers in vitro and could differentiate into embryonic tissues in vivo. NT embryos from early passage R1 donor cells were able to form full term developed pups, whereas those from late passage R1 ES donor cells lost the potential for reprogramming that is essential for live birth. We subsequently established sequential NT-R1-ESC lines that were developed from NT blastocyst of late passage R1 ESC donors. However, these NT-R1-ESC lines, when used as nuclear transfer donors at their early passages, failed to result in live pups. This indicates that the therapeutic cloning process using sequential NT-ESCs may not rescue the developmental deficiencies that resided in previous donor generations.

Cell Research advance online publication 9 January 2007; doi: 10.1038/sj.cr.7310139

Keywords: nuclear transfer, therapeutic cloning, embryonic stem cells