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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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.
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). https://doi.org/10.1038/nature11435
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