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.
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Abbreviations
- NT:
-
nuclear transfer
- ntES cells:
-
embryonic stem cells derived from somatic cells by NT
- EBs:
-
embryoid bodies
- SCNT:
-
somatic cell nuclear transfer
- FBS:
-
fetal bovine serum
References
Solter D, J Gearhart . Putting stem cells to work. Science 1999; 283:1468–70.
Lanza RP, JB Cibelli, MD West . Human therapeutic cloning. Nature Med 1999; 5:975–7.
Colman A, A Kind . Therapeutic cloning: concepts and practicalities. Trends Biotechnol 2000; 18:192–6.
Cibelli JB, SL Stice, PJ Golueke, et al. Transgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells. Nature Biotechnol 1998; 16:642–6.
Munsie MJ, AE Michalska, CM O'Brien, AO Trounson, MF Pera, PS Mountford . Isolation of pluripotent embryonic stem cells from reprogrammed adult mouse somatic cell nuclei. Curr Biol 2000; 10:989–92.
Kawase E, Y Yamazaki, T Yagi, R Yanagimachi, RA Pedersen . Mouse embryonic stem (ES) cell lines established from neuronal cell-derived cloned blastocysts. Genesis 2000; 28:156–63.
Wakayama T, V Tabar, I Rodriguez, AC Perry, L Studer, P Mombaerts . Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer. Science 2001; 292:740–3.
Simerly C, T Cominko, C Navara, et al. Molecular correlates of primate nuclear transfer failures. Science 2003; 300:297.
Dominko T, M Mitalipova, B Haley, et al. Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species. Biol Reprod 1999; 60:1496–1502.
Yang X, S Jiang, A Kovás, RH Foote . Nuclear totipotency of cultured rabbit morulae to support full-term development following nuclear transfer. Biol Reprod 1992; 47:636–43.
Chesné P, PG Adenot, C Viglietta, M Baratte, L Boulanger, JP Renard . Cloned rabbits produced by nuclear transfer from adult somatic cells. Nature Biotechnology 2002; 20:366–9.
Chen DY, QY Sun, JL Liu, et al. Dedifferentiation of giant panda (Ailuropoda melanoleuca) somatic nuclei in rabbit ooplasm and early development of the reconstructed egg. Science in China (Series C) 1999; 29:24–330.
Thomson JA, J Itskovitz-Eldor, SS Shapiro, et al. Embryonic stem cell lines derived from human blastocysts. Science 1998; 282:1145–7.
Itskovitz-Eldor J, M Schuldiner, D Karsenti, et al. Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers. Mol Med 2000; 6:88–95.
Reubinoff BE, MF Pera, CY Fong, A Trounson, A Bongso . Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nature Biotechnol 2000; 18:399–404.
Shamblott MJ, J Axelman, JW Littlefield, et al. Human embryonic germ cell derivatives express a broad range of developmentally distinct markers and proliferate extensively in vitro. Proc Natl Acad Sci USA 2001; 98:113–8.
Sambrook J, DW Russell . Molecular Cloning: a Laboratory Manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York). 2001.
Brüstle O, K Choudhary, K Karram, et al. Chimeric brains generated by intraventricular transplantation of fetal human brain cells into embryonic rats. Nat Biotechnol 1998; 16:1040–4.
Rubin CM, CM Houck, PL Deininger, T Friedmann, CW Schmid . Partial nucleotide sequence of the 300-nucleotide interspersed repeated human DNA sequences. Nature 1980; 284:372–4.
Van de Corput MP, JM van den Ouweland, RW Dirks, LM Hart, GJ Bruining, JA Maassen, AK Raap . Detection of mitochondrial DNA deletions in human skin fibroblasts of patients with Pearson's syndrome by two-color fluorescence in situ hybridization. J Histochem Cytochem 1997; 45:55–61.
Dunham I, C Lengauer, T Cremer, T Featherstone . Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction. Hum Genet 1992; 88:457–62.
Wakayama T, R Yanagimachi . Mouse cloning with nucleus donor cells of different age and type. Mol Reprod Dev 2001; 58:376–83.
Kubota C, H Yamakuchi, J Todoroki, et al. Six cloned calves produced from adult fibroblast cells after long-term culture. Proc Natl Acad Sci USA 2000; 97:990–5.
Wang WH, L Meng, RJ Hackett, R Odenbourg, DL Keefe . The spindle observation and its relationship with fertilization after intracytoplasmic sperm injection in living human oocytes. Fertility and Sterility 2001; 75:348–53.
Pera MF, B Reubinoff, A Trounson . Human embryonic stem cells. J Cell Sci 2000; 113:5–10.
Kim NW, MA Piatyszek, KR Prowse, et al. Specific association of human telomerase activity with immortal cells and cancer. Science 1994; 266:2011–5.
Doetschman TC, H Eistetter, M Katz, W Schmidt, R Kemler . The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morph 1985; 87:27–45.
Rathjen PD, J Lake, LM Whyatt, MD Bettess, J Rathjen . Properties and uses of embryonic stem cells: prospects for application to human biology and gene therapy. Reprod Fertil Dev 1998; 10:31–47.
Brivanlou AH, FH Gage, R Jaenisch, T Jessell, D Melton, J Rossant . Setting standards for human embryonic stem cells. Science 2003; 300:913–6.
Odorico JS, DS Kaufman, JA Thomson . Multilineage differentiation from human embryonic stem cell lines. Stem Cells 2001; 19:193–204.
Acknowledgements
We are grateful to Drs. M Brown, JL Goldstein, DL Garbers and R Hammer for critical reading of the manuscript. We thank You Ming ZHU, Zheng Ya TANG, Wen Jun DING, Chao PENG, Min Jie NI, Yang WANG for their excellent technical support and Dr. Lan Dian HU for her assistance in DNA microsatellite analysis. Also, we thank Dr. Li Jun ZHANG for karyotyping and Dr. Qiang Su GUO for assistance in confocal microscopic analysis.
This work was supported by grants from the Major State Basic Research Development Program of China. (No. 001CB5099), the National High Technology Research and Development Program of China (No. 2001AA216121), National Natural Science Foundation of China (No. 30040003), Projects of Shanghai Science & Technology Development Foundation (No. 99DJ14002, 00DJ14033, 01DJ14003), the Chinese Academy of Sciences (No. KSCX-2-3-08), Shanghai Municipal Education Commission and by Shanghai Second Medical University.
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CHEN, Y., HE, Z., LIU, A. et al. Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes. Cell Res 13, 251–263 (2003). https://doi.org/10.1038/sj.cr.7290170
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DOI: https://doi.org/10.1038/sj.cr.7290170
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