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Androgenetic haploid embryonic stem cells produce live transgenic mice


Haploids and double haploids are important resources for studying recessive traits and have large impacts on crop breeding1, but natural haploids are rare in animals. Mammalian haploids are restricted to germline cells and are occasionally found in tumours with massive chromosome loss2,3. Recent success in establishing haploid embryonic stem (ES) cells in medaka fish4 and mice5,6 raised the possibility of using engineered mammalian haploid cells in genetic studies. However, the availability and functional characterization of mammalian haploid ES cells are still limited. Here we show that mouse androgenetic haploid ES (ahES) cell lines can be established by transferring sperm into an enucleated oocyte. The ahES cells maintain haploidy and stable growth over 30 passages, express pluripotent markers, possess the ability to differentiate into all three germ layers in vitro and in vivo, and contribute to germlines of chimaeras when injected into blastocysts. Although epigenetically distinct from sperm cells, the ahES cells can produce viable and fertile progenies after intracytoplasmic injection into mature oocytes. The oocyte-injection procedure can also produce viable transgenic mice from genetically engineered ahES cells. Our findings show the developmental pluripotency of androgenentic haploids and provide a new tool to quickly produce genetic models for recessive traits. They may also shed new light on assisted reproduction.

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Figure 1: Generation of androgenetic haploid ES cells.
Figure 2: The pluripotency of the ahES cells.
Figure 3: Generation of ICAI offspring and transgenic mice.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

CGH data and gene-expression data are deposited at the Gene Expression Omnibus under accession numbers GSE39390 and GSE39391, respectively.


  1. 1

    Germanà, M. A. Gametic embryogenesis and haploid technology as valuable support to plant breeding. Plant Cell Rep. 30, 839–857 (2011)

    Article  Google Scholar 

  2. 2

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

    ADS  CAS  Article  Google Scholar 

  3. 3

    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 

  4. 4

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

    ADS  CAS  Article  Google Scholar 

  5. 5

    Leeb, M. & Wutz, A. Derivation of haploid embryonic stem cells from mouse embryos. Nature 479, 131–134 (2011)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Elling, U. et al. Forward and reverse genetics through derivation of haploid mouse embryonic stem cells. Cell Stem Cell 9, 563–574 (2011)

    CAS  Article  Google Scholar 

  7. 7

    Zhou, Q. et al. Generation of fertile cloned rats by regulating oocyte activation. Science 302, 1179 (2003)

    CAS  Article  Google Scholar 

  8. 8

    Yanagimachi, R., Wakayama, T., Perry, A. C. F., Zuccotti, M. & Johnson, K. R. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–374 (1998)

    ADS  Article  Google Scholar 

  9. 9

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

    ADS  CAS  Article  Google Scholar 

  10. 10

    Yoshimizu, T. et al. Germline-specific expression of the Oct-4/green fluorescent protein (GFP) transgene in mice. Dev. Growth Differ. 41, 675–684 (1999)

    CAS  Article  Google Scholar 

  11. 11

    Tarkowki, A. K. In vitro development of haploid mouse embryos produced by bisection of one-cell fertilized eggs. J. Embryol. Exp. Morphol. 38, 187–202 (1977)

    CAS  PubMed  Google Scholar 

  12. 12

    Latham, K. E., Patel, B., Bautista, F. D. & Hawes, S. M. Effects of X chromosome number and parental origin on X-linked gene expression in preimplantation mouse embryos. Biol. Reprod. 63, 64–73 (2000)

    CAS  Article  Google Scholar 

  13. 13

    Yang, H. et al. Generation of genetically modified mice by oocyte injection of androgenetic haploid embryonic stem cells. Cell 149, 605–617 (2012)

    CAS  Article  Google Scholar 

  14. 14

    Nichols, J. & Smith, A. Naive and primed pluripotent states. Cell Stem Cell 4, 487–492 (2009)

    CAS  Article  Google Scholar 

  15. 15

    Brons, I. G. et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448, 191–195 (2007)

    ADS  CAS  Article  Google Scholar 

  16. 16

    Tesar, P. J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196–199 (2007)

    ADS  CAS  Article  Google Scholar 

  17. 17

    Maruotti, J. et al. Nuclear transfer-derived epiblast stem cells are transcriptionally and epigenetically distinguishable from their fertilized-derived counterparts. Stem Cells 28, 743–752 (2010)

    CAS  Article  Google Scholar 

  18. 18

    Kimura, Y. & Yanagimachi, R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development 121, 2397–2405 (1995)

    CAS  PubMed  Google Scholar 

  19. 19

    Loren, J. & Lacham-Kaplan, O. The employment of strontium to activate mouse oocytes: effects on spermatid-injection outcome. Reproduction 131, 259–267 (2006)

    CAS  Article  Google Scholar 

  20. 20

    Mayer, W., Niveleau, A., Walter, J., Fundele, R. & Haaf, T. Demethylation of the zygotic paternal genome. Nature 403, 501–502 (2000)

    ADS  CAS  Article  Google Scholar 

  21. 21

    Santos, F., Hendrich, B., Reik, W. & Dean, W. Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev. Biol. 241, 172–182 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Surani, M. A., Barton, S. C. & Norris, M. L. Development of reconstituted mouse eggs suggests imprinting of the genome during gametogenesis. Nature 308, 548–550 (1984)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Surani, M. A. et al. Genome imprinting and development in the mouse. Dev. (Suppl.). 89–98 (1990)

  24. 24

    McGrath, J. & Solter, D. Completion of mouse embryogenesis requires both the maternal and paternal genomes. Cell 37, 179–183 (1984)

    CAS  Article  Google Scholar 

  25. 25

    Tsai, T. F., Jiang, Y. H., Bressler, J., Armstrong, D. & Beaudet, A. L. Paternal deletion from Snrpn to Ube3a in the mouse causes hypotonia, growth retardation and partial lethality and provides evidence for a gene contributing to Prader-Willi syndrome. Hum. Mol. Genet. 8, 1357–1364 (1999)

    CAS  Article  Google Scholar 

  26. 26

    Kono, T. et al. Birth of parthenogenetic mice that can develop to adulthood. Nature 428, 860–864 (2004)

    ADS  CAS  Article  Google Scholar 

  27. 27

    Tachibana, M. et al. Generation of chimeric rhesus monkeys. Cell 148, 285–295 (2012)

    CAS  Article  Google Scholar 

  28. 28

    Mann, J. R., Gadi, I., Harbison, M. L., Abbondanzo, S. J. & Stewart, C. L. Androgenetic mouse embryonic stem cells are pluripotent and cause skeletal defects in chimeras: implications for genetic imprinting. Cell 62, 251–260 (1990)

    CAS  Article  Google Scholar 

  29. 29

    Zhao, X. Y. et al. Viable fertile mice generated from fully pluripotent iPS cells derived from adult somatic cells. Stem Cell Rev. 6, 390–397 (2010)

    Article  Google Scholar 

  30. 30

    Ying, Q. L. & Smith, A. G. Defined conditions for neural commitment and differentiation. Methods Enzymol. 365, 327–341 (2003)

    CAS  Article  Google Scholar 

  31. 31

    Sheng, C. et al. Direct reprogramming of Sertoli cells into multipotent neural stem cells by defined factors. Cell Res. 22, 208–218 (2012)

    CAS  Article  Google Scholar 

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We thank all members of the Group of Reproductive Engineering for discussion and help. This study was supported by a grant from the National Science Foundation of China 90919060 (to Q.Z.) and the China National Basic Research Program 2012CBA01300 (to Q.Z.), and a grant from the Strategic Priority Research Program of the Chinese Academy of Sciences XDA01020100 (to Q.Z.). We thank Fluidigm Corporation for their support in the utilization of BioMark HD system. We thank Eppendorf and Leica for supporting the facility.

Author information




Q.Z. and X.-Y.Z. designed the experiments;; W.L., L.S., H.W., M.D., M.W., L.S., C.F., T.L., X.L., L.W., Q.-Y.Z. and C.S. performed experiments; Q.Z, X.-Y.Z, X.-J.W., G.-Z.L., W.L., H.-J.W. and L.L. analysed data; Q.Z., X.-Y.Z., Z.L., and L.W..supervised experiments; W.L., L.S., H.W. and M.D. contributed to part of the Methods. Q.Z., X.-Y.Z., X.-J.W. and W.L. wrote the paper.

Corresponding authors

Correspondence to Xiao-Yang Zhao or Qi Zhou.

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The authors declare no competing financial interests.

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Li, W., Shuai, L., Wan, H. et al. Androgenetic haploid embryonic stem cells produce live transgenic mice. Nature 490, 407–411 (2012).

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