1. ¹Center for Developmental Biology, Shanghai Second Medical University, Shanghai 200092, China.
2. ²Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 1665 Kong Jiang Road, Shanghai 200092, China.
3. ³Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510120, China.
4. ⁴State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 19 Zhong-guan Cun Road, Haidian District, Beijing 100080, China.
5. ⁵Health Science Center, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China.
6. ⁶National Laboratory of Medical Genetics, Central South University, 88 Xiangya Road, Changsha 410078, China.
7. ⁷Xinhua Hospital, Shanghai Second Medical University, 1665 Kong Jiang Road, Shanghai 200092, China

Correspondence: Hui Zhen SHENG, Center for Developmental Biology, 1665 Kong Jiang Road, Xinhua Hospital, Shanghai Second Medical University, Shanghai, 200092 China. Telephone and fax number: 0086-21-55570017. E-mail: 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

Received 4 August 2003; Revised 11 August 2003; Accepted 13 August 2003.

Abstract

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.