Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Human embryonic stem cell lines derived from single blastomeres

Nature volume 444, pages 481–485 (2006)Cite this article

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.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

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.

References

  1. Lanza, R. et al. (eds) Essentials of Stem Cell Biology (Elsevier/Academic, San Diego, 2006)

  2. Thomson, J. A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147 (1998)

    Article  ADS  CAS  Google Scholar 

  3. Cowan, C. A. et al. Derivation of embryonic stem-cell lines from human blastocysts. N. Engl. J. Med. 350, 1353–1356 (2004)

    Article  CAS  Google Scholar 

  4. Strelchenko, N. et al. Morula-derived human embryonic stem cells. Reprod. BioMed. 9, 623–629 (2004)

    Article  Google Scholar 

  5. Chung, Y. et al. Embryonic and extraembryonic stem cell lines derived from single mouse blastomeres. Nature 439, 216–219 (2006)

    Article  ADS  CAS  Google Scholar 

  6. Veeck, L. L. et al. An Atlas of Human Gametes and Conceptuses (Parthenon, New York, 1999)

    Book  Google Scholar 

  7. Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R. (eds) Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2002)

  8. Klimanskaya, I. & McMahon, J. in Handbook of Stem Cells Vol. 1 Embryonic Stem Cells (eds Lanza, R. et al.) 437–449 (Elsevier/Academic, San Diego, 2004)

    Google Scholar 

  9. Klimanskaya, I. et al. Human embryonic stem cells derived without feeder cells. Lancet 361, 1496–1501 (2005)

    Google Scholar 

  10. Klimanskaya, I. et al. Derivation and comparative assessment of retinal pigment epithelium from human embryonic stem cells using transcriptomics. Clon. Stem Cells 6, 217–245 (2004)

    Article  CAS  Google Scholar 

  11. Lanzendorf, S. E. et al. Use of human gametes obtained from anonymous donors for the production of human embryonic stem cell lines. Fertil. Steril. 76, 132–137 (2001)

    Article  CAS  Google Scholar 

  12. Mitalipova, M. et al. Human embryonic stem cell lines derived from discarded embryos. Stem Cells 21, 521–526 (2003)

    Article  CAS  Google Scholar 

  13. Weber, D. J. Considerations for clinical uses of therapies derived from stem cells. In Methods in Enzymology Vol. 420 Stem Cell Tools and Therapeutic Applications (eds Klimanskaya, I. & Lanza, R.) (Elsevier, New York, in the press)

  14. Richards, M., Fong, C. Y., Chan, W. K., Wong, P. C. & Bongso, A. Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells. Nature Biotechnol. 20, 933–936 (2002)

    Article  CAS  Google Scholar 

  15. Ludwig, T. E. et al. Derivation of human embryonic stem cells in defined conditions. Nature Biotechnol. 24, 185–187 (2006)

    Article  CAS  Google Scholar 

  16. Young, H., Xu, C. & Carpenter, M. K. in Essentials of Stem Cell Biology (eds Lanza, R. et al.) 317–324 (Elsevier/Academic, San Diego, 2006)

    Google Scholar 

  17. Deb, K., Sivaguru, M., Yong, H. Y. & Roberts, R. M. Cdx2 gene expression and trophectoderm lineage specification in mouse embryos. Science 311, 992–996 (2006)

    Article  ADS  CAS  Google Scholar 

  18. Piotrowska-Nitsche, K., Perea-Gomez, A., Haraguchi, S. & Zernicka-Goetz, Z. Four-cell stage mouse blastomeres have different developmental properties. Development 132, 479–490 (2005)

    Article  CAS  Google Scholar 

  19. Hansis, C., Tang, Y., Grifo, J. A. & Krey, L. C. Analysis of Oct-4 expression and ploidy in individual human blastomeres. Mol. Hum. Reprod. 7, 155–161 (2001)

    Article  CAS  Google Scholar 

  20. Geber, S., Winston, R. M. L. & Handyside, A. H. Proliferation of blastomeres from cleavage stage human embryos in vitro: an alternative to blastocyst biopsy for preimplantation diagnosis. Hum. Reprod. 10, 1492–1496 (1995)

    Article  CAS  Google Scholar 

  21. Sills, E. S., Takeuchi, T., Tanaka, N., Queenie, V. N. & Palermo, G. D. Identification and isolation of embryonic stem cells in reproductive endocrinology: theoretical protocols for conservation of human embryos derived from in vitro fertilization. Theor. Biol. Med. Model 2, 25 (2005)

    Article  Google Scholar 

  22. Handyside, A. H. et al. Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature 344, 768–770 (1990)

    Article  ADS  CAS  Google Scholar 

  23. Magli, M. C. et al. Cryopreservation of biopsied embryos at the blastocyst stage. Hum. Reprod. doi:10.1093/humrep/del228 (published online 12 June 2006)

  24. Hardy, K., Martin, K. L., Leese, H., Winston, R. M. & Handyside, A. Human preimplantation development in vitro is not adversely affected by biopsy at the 8-cell stage. Hum. Reprod. 5, 708–714 (1990)

    Article  CAS  Google Scholar 

  25. Staessen, C. et al. Comparison of blastocyst transfer with or without preimplantation genetic diagnosis for aneuploidy screening in couples with advanced maternal age: a prospective randomized controlled trial. Hum. Reprod. 19, 2849–2858 (2004)

    Article  Google Scholar 

  26. Fredsted, T. & Villesen, P. Fast and reliable sexing of prosimian and human DNA. Am. J. Pathol. 64, 345–350 (2004)

    CAS  Google Scholar 

  27. Koopman, P., Gubbay, J., Vivian, N., Goodfellow, P. & Lovell-Badge, R. Male development of chromosomally female mice transgenic for Sry. Nature 351, 117–121 (1991)

    Article  ADS  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  1. Irina Klimanskaya and Young Chung: *These authors contributed equally to this work

Authors and Affiliations

  1. Advanced Cell Technology, 381 Plantation Street, Massachusetts, 01605, Worcester, USA

    Irina Klimanskaya, Young Chung, Sandy Becker, Shi-Jiang Lu & Robert Lanza

Authors
  1. Irina Klimanskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Young Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sandy Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shi-Jiang Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert Lanza
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Robert Lanza.

Ethics declarations

Competing interests

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)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature05142

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing