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Human embryonic stem cell lines derived from single blastomeres

A Corrigendum to this article was published on 15 March 2007

An Addendum to this article was published on 23 November 2006

Abstract

The derivation of human embryonic stem (hES) cells currently requires the destruction of ex utero embryos1,2,3,4. A previous study in mice indicates that it might be possible to generate embryonic stem (ES) cells using a single-cell biopsy similar to that used in preimplantation genetic diagnosis (PGD), which does not interfere with the embryo’s developmental potential5. By growing the single blastomere overnight, the resulting cells could be used for both genetic testing and stem cell derivation without affecting the clinical outcome of the procedure. Here we report a series of ten separate experiments demonstrating that hES cells can be derived from single blastomeres. In this proof-of-principle study, multiple biopsies were taken from each embryo using micromanipulation techniques and none of the biopsied embryos were allowed to develop in culture. Nineteen ES-cell-like outgrowths and two stable hES cell lines were obtained. The latter hES cell lines maintained undifferentiated proliferation for more than eight months, and showed normal karyotype and expression of markers of pluripotency, including Oct-4, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, nanog and alkaline phosphatase. These cells retained the potential to form derivatives of all three embryonic germ layers both in vitro and in teratomas. The ability to create new stem cell lines and therapies without destroying embryos would address the ethical concerns of many, and allow the generation of matched tissue for children and siblings born from transferred PGD embryos.

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Figure 1: Derivation of hES cells from single blastomeres.
Figure 2: Characterization of hES cells from single blastomeres.
Figure 3: Microsatellite and PCR analysis of single-blastomere-derived hES cells versus the cell line (WA01) used for co-culture.
Figure 4: In vitro differentiation of single-blastomere-derived hES cells into all three germ layers.

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Acknowledgements

We thank T. Li and A. Feng for their assistance with hES cell culture and differentiation, S. Agarwal, J. Shepard and T. Lin for RT–PCR, J. Johnson and L. Meisner for karyotype analysis, and J. Staruk and L. McGill for histological analysis.

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Correspondence to Robert Lanza.

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The hES cell lines generated in this study will be made available to investigators under a material transfer agreement (an application has been submitted for deposition of the hES cell lines into the UK Stem Cell Bank). Reprints and permissions information is available at www.nature.com/reprints. The authors declare competing financial interests: details accompany the paper at www.nature.com/nature.

Supplementary information

Supplementary Figure 1

Original DNA gels. a–c, uncropped gels for Figure 4 a-c, respectively. d, uncropped gel for Figure 3i. (JPG 24 kb)

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Klimanskaya, I., Chung, Y., Becker, S. et al. Human embryonic stem cell lines derived from single blastomeres. Nature 444, 481–485 (2006). https://doi.org/10.1038/nature05142

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