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.

  • Letter
  • Published:

Derivation of haploid embryonic stem cells from mouse embryos

Nature volume 479, pages 131–134 (2011)Cite this article

Subjects

Abstract

Most animals are diploid, but haploid-only and male-haploid (such as honeybee and ant) species have been described1. The diploid genomes of complex organisms limit genetic approaches in biomedical model species such as mice. To overcome this problem, experimental induction of haploidy has been used in fish2,3. Haploid development in zebrafish has been applied for genetic screening2. Recently, haploid pluripotent cell lines from medaka fish (Oryzias latipes) have also been established3. In contrast, haploidy seems less compatible with development in mammals4,5. Although haploid cells have been observed in egg cylinder stage parthenogenetic mouse embryos6, most cells in surviving embryos become diploid. Here we describe haploid mouse embryonic stem cells and show their application in forward genetic screening.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Derivation of haploid ES cells.
Figure 2: Expression analysis of haploid ES cells.
Figure 3: Developmental potential of haploid ES cells.

Similar content being viewed by others

Accession codes

Primary accessions

Gene Expression Omnibus

References

  1. Otto, S. P. & Jarne, P. Evolution. Haploids—hapless or happening? Science 292, 2441–2443 (2001)

    Article  CAS  Google Scholar 

  2. Wiellette, E. et al. Combined haploid and insertional mutation screen in the zebrafish. Genesis 40, 231–240 (2004)

    Article  CAS  Google Scholar 

  3. Yi, M., Hong, N. & Hong, Y. Generation of medaka fish haploid embryonic stem cells. Science 326, 430–433 (2009)

    Article  ADS  CAS  Google Scholar 

  4. Kaufman, M. H., Robertson, E. J., Handyside, A. H. & Evans, M. J. Establishment of pluripotential cell lines from haploid mouse embryos. J. Embryol. Exp. Morphol. 73, 249–261 (1983)

    CAS  PubMed  Google Scholar 

  5. Latham, K. E., Akutsu, H., Patel, B. & Yanagimachi, R. Comparison of gene expression during preimplantation development between diploid and haploid mouse embryos. Biol. Reprod. 67, 386–392 (2002)

    Article  CAS  Google Scholar 

  6. Kaufman, M. H. Chromosome analysis of early postimplantation presumptive haploid parthenogenetic mouse embryos. J. Embryol. Exp. Morphol. 45, 85–91 (1978)

    CAS  PubMed  Google Scholar 

  7. Ying, Q. L. et al. The ground state of embryonic stem cell self-renewal. Nature 453, 519–523 (2008)

    Article  ADS  CAS  Google Scholar 

  8. Nichols, J. et al. Validated germline-competent embryonic stem cell lines from nonobese diabetic mice. Nature Med. 15, 814–818 (2009)

    Article  CAS  Google Scholar 

  9. Buehr, M. et al. Capture of authentic embryonic stem cells from rat blastocysts. Cell 135, 1287–1298 (2008)

    Article  CAS  Google Scholar 

  10. Nichols, J., Silva, J., Roode, M. & Smith, A. Suppression of Erk signalling promotes ground state pluripotency in the mouse embryo. Development 136, 3215–3222 (2009)

    Article  CAS  Google Scholar 

  11. Nichols, J. & Smith, A. The origin and identity of embryonic stem cells. Development 138, 3–8 (2011)

    Article  CAS  Google Scholar 

  12. Cutler, G., Marshall, L. A., Chin, N., Baribault, H. & Kassner, P. D. Significant gene content variation characterizes the genomes of inbred mouse strains. Genome Res. 17, 1743–1754 (2007)

    Article  CAS  Google Scholar 

  13. Pollard, S. M., Benchoua, A. & Lowell, S. Neural stem cells, neurons, and glia. Methods Enzymol. 418, 151–169 (2006)

    Article  CAS  Google Scholar 

  14. Li, M. A., Pettitt, S. J., Yusa, K. & Bradley, A. Genome-wide forward genetic screens in mouse ES cells. Methods Enzymol. 477, 217–242 (2010)

    Article  CAS  Google Scholar 

  15. Cadinanos, J. & Bradley, A. Generation of an inducible and optimized piggyBac transposon system. Nucleic Acids Res. 35, e87 (2007)

    Article  Google Scholar 

  16. Mikkers, H. et al. High-throughput retroviral tagging to identify components of specific signaling pathways in cancer. Nature Genet. 32, 153–159 (2002)

    Article  CAS  Google Scholar 

  17. Mai, Q. et al. Derivation of human embryonic stem cell lines from parthenogenetic blastocysts. Cell Res. 17, 1008–1019 (2007)

    Article  ADS  CAS  Google Scholar 

  18. Revazova, E. S. et al. Patient-specific stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells 9, 432–449 (2007)

    Article  CAS  Google Scholar 

  19. Inoue, K. et al. Impeding Xist expression from the active X chromosome improves mouse somatic cell nuclear transfer. Science 330, 496–499 (2010)

    Article  ADS  CAS  Google Scholar 

  20. Sukov, W. R. et al. Nearly identical near-haploid karyotype in a peritoneal mesothelioma and a retroperitoneal malignant peripheral nerve sheath tumor. Cancer Genet. Cytogenet. 202, 123–128 (2010)

    Article  CAS  Google Scholar 

  21. Kotecki, M., Reddy, P. S. & Cochran, B. H. Isolation and characterization of a near-haploid human cell line. Exp. Cell Res. 252, 273–280 (1999)

    Article  CAS  Google Scholar 

  22. Carette, J. E. et al. Haploid genetic screens in human cells identify host factors used by pathogens. Science 326, 1231–1235 (2009)

    Article  ADS  CAS  Google Scholar 

  23. Jiang, H. et al. Activation of paternally expressed imprinted genes in newly derived germline-competent mouse parthenogenetic embryonic stem cell lines. Cell Res. 17, 792–803 (2007)

    Article  CAS  Google Scholar 

  24. Kishigami, S. & Wakayama, T. Efficient strontium-induced activation of mouse oocytes in standard culture media by chelating calcium. J. Reprod. Dev. 53, 1207–1215 (2007)

    Article  CAS  Google Scholar 

  25. Guo, G., Wang, W. & Bradley, A. Mismatch repair genes identified using genetic screens in Blm-deficient embryonic stem cells. Nature 429, 891–895 (2004)

    Article  ADS  CAS  Google Scholar 

  26. Savarese, F., Flahndorfer, K., Jaenisch, R., Busslinger, M. & Wutz, A. Hematopoietic precursor cells transiently reestablish permissiveness for X inactivation. Mol. Cell. Biol. 26, 7167–7177 (2006)

    Article  CAS  Google Scholar 

  27. Wutz, A. & Jaenisch, R. A shift from reversible to irreversible X inactivation is triggered during ES cell differentiation. Mol. Cell 5, 695–705 (2000)

    Article  CAS  Google Scholar 

  28. Leeb, M. et al. Polycomb complexes act redundantly to repress genomic repeats and genes. Genes Dev. 24, 265–276 (2010)

    Article  CAS  Google Scholar 

  29. Shen, X. et al. EZH1 mediates methylation on histone H3 lysine 27 and complements EZH2 in maintaining stem cell identity and executing pluripotency. Mol. Cell 32, 491–502 (2008)

    Article  MathSciNet  CAS  Google Scholar 

  30. Kong, J., Zhu, F., Stalker, J. & Adams, D. J. iMapper: a web application for the automated analysis and mapping of insertional mutagenesis sequence data against Ensembl genomes. Bioinformatics 24, 2923–2925 (2008)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank A. Smith and J. Nichols for critical discussion; K. Jones for advice on cell culture; S. Dietmann for bioinformatics support; and R. Walker for cell sorting. We would also like to thank B. Mansfield and C.-E. Dumeau for their help, and the BSU team at the centre for maintaining the mouse colony. This work was supported by a Wellcome Trust Senior Research Fellowship to A.W. (grant reference 087530/Z/08/A) and an EMBO Long Term Fellowship to M.L.

Author information

Authors and Affiliations

  1. Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

    Martin Leeb & Anton Wutz

Authors
  1. Martin Leeb
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anton Wutz
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.L. performed the experiments, analysed the data and wrote the manuscript. A.W. performed some experiments, wrote the paper and supervised the study.

Corresponding author

Correspondence to Anton Wutz.

Ethics declarations

Competing interests

A patent application covering haploid embryonic stem cells has been filed.

Additional information

Gene expression and CGH data sets can be accessed as the GEO reference series GSE30879 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE30879). This series includes the GSE30744 (Expression analysis of haploid and diploid ES cells in 2i medium) and the GSE30749 (CGH analysis of haploid ES cells) data sets.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-8 with legends and Supplementary Tables 1-2. (PDF 1822 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leeb, M., Wutz, A. Derivation of haploid embryonic stem cells from mouse embryos. Nature 479, 131–134 (2011). https://doi.org/10.1038/nature10448

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10448

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