REGULAR ARTICLES

Cell Research (2003); 13(4):251-263

Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes

Ying CHEN^{1, 2}, Zhi Xu HE³, Ailian LIU^{1, 2}, Kai WANG^{1, 2}, Wen Wei MAO^{1, 2}, Jian Xin CHU^{1, 2}, Yong LU^{1, 2}, Zheng Fu FANG^{1, 2}, Ying Tang SHI^{1, 2}, Qing Zhang YANG^{1, 2}, Da Yuan CHEN⁴, Min Kang WANG⁴, Jin Song LI⁴, Shao Liang HUANG³, Xiang Yin KONG⁵, Yao Zhou SHI⁵, Zhi Qiang WANG⁵, Jia Hui XIA⁶, Zhi Gao LONG⁶, Zhi Gang XUE⁶, Wen Xiang DING⁷, Hui Zhen SHENG^{1, 2, *}

Correspondence: Dr. Hui Zhen SHENG Center for Developmental Biology, 1665 Kong Jiang Road, Xinhua Hospital, Shanghai Second Medical University, Shanghai, 200092 China. 0086-21-55570017 (phone & fax) hzsheng@sh163a.sta.net.cn Dr. Hui Zhen SHENG is also an adjunct faculty member in the Department of Pharmacology/Cecil H. & Ida Green Center for Reproductive Biology Sciences, The University of Texas Southwestern Medical Center, Dallas, TX, USA

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To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well as mixed cell populations that express markers representative of all three germ layers. Thus, ntES cells derived from human somatic cells by NT to rabbit eggs retain phenotypes similar to those of conventional human ES cells, including the ability to undergo multilineage cellular differentiation.

Key words: nuclear transfer (NT), somatic cell nuclear transfer (SCNT), embryonic stem cells (ES cell), therapeutic cloning, rabbit oocyte.