Skip to main content

Thank you for visiting 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.

Mitochondrial gene replacement in primate offspring and embryonic stem cells

A Corrigendum to this article was published on 10 December 2014


Mitochondria are found in all eukaryotic cells and contain their own genome (mitochondrial DNA or mtDNA). Unlike the nuclear genome, which is derived from both the egg and sperm at fertilization, the mtDNA in the embryo is derived almost exclusively from the egg; that is, it is of maternal origin. Mutations in mtDNA contribute to a diverse range of currently incurable human diseases and disorders. To establish preclinical models for new therapeutic approaches, we demonstrate here that the mitochondrial genome can be efficiently replaced in mature non-human primate oocytes (Macaca mulatta) by spindle–chromosomal complex transfer from one egg to an enucleated, mitochondrial-replete egg. The reconstructed oocytes with the mitochondrial replacement were capable of supporting normal fertilization, embryo development and produced healthy offspring. Genetic analysis confirmed that nuclear DNA in the three infants born so far originated from the spindle donors whereas mtDNA came from the cytoplast donors. No contribution of spindle donor mtDNA was detected in offspring. Spindle replacement is shown here as an efficient protocol replacing the full complement of mitochondria in newly generated embryonic stem cell lines. This approach may offer a reproductive option to prevent mtDNA disease transmission in affected families.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Spindle–chromosomal complex transfer and meiotic analysis of reconstructed monkey oocytes.
Figure 2: Mito and Tracker, the first primates to be produced by spindle–chromosomal complex transfer (ST) into enucleated oocytes followed by fertilization and embryo transfer.
Figure 3: MtDNA analysis in ST offspring.


  1. Wallace, D. C. et al. Mitochondrial DNA mutation associated with Leber’s hereditary optic neuropathy. Science 242, 1427–1430 (1988)

    Article  ADS  CAS  Google Scholar 

  2. Holt, I. J., Harding, A. E. & Morgan-Hughes, J. A. Deletions of muscle mitochondrial DNA in patients with mitochondrial myopathies. Nature 331, 717–719 (1988)

    Article  ADS  CAS  Google Scholar 

  3. Zeviani, M. et al. Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology 38, 1339–1346 (1988)

    Article  CAS  Google Scholar 

  4. Solano, A., Playan, A., Lopez-Perez, M. J. & Montoya, J. Genetic diseases of the mitochondrial DNA in humans [in Spanish]. Salud Publica Mex. 43, 151–161 (2001)

    Article  CAS  Google Scholar 

  5. Keating, D. J. Mitochondrial dysfunction, oxidative stress, regulation of exocytosis and their relevance to neurodegenerative diseases. J. Neurochem. 104, 298–305 (2008)

    CAS  PubMed  Google Scholar 

  6. Lin, M. T. & Beal, M. F. Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443, 787–795 (2006)

    Article  ADS  CAS  Google Scholar 

  7. Reeve, A. K., Krishnan, K. J. & Turnbull, D. Mitochondrial DNA mutations in disease, aging, and neurodegeneration. Ann. NY Acad. Sci. 1147, 21–29 (2008)

    Article  ADS  CAS  Google Scholar 

  8. Trushina, E. & McMurray, C. T. Oxidative stress and mitochondrial dysfunction in neurodegenerative diseases. Neuroscience 145, 1233–1248 (2007)

    Article  CAS  Google Scholar 

  9. Brandon, M., Baldi, P. & Wallace, D. C. Mitochondrial mutations in cancer. Oncogene 25, 4647–4662 (2006)

    Article  CAS  Google Scholar 

  10. Sutovsky, P. et al. Ubiquitin tag for sperm mitochondria. Nature 402, 371–372 (1999)

    Article  ADS  CAS  Google Scholar 

  11. Majamaa, K. et al. Epidemiology of A3243G, the mutation for mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes: prevalence of the mutation in an adult population. Am. J. Hum. Genet. 63, 447–454 (1998)

    Article  CAS  Google Scholar 

  12. Taylor, R. W. & Turnbull, D. M. Mitochondrial DNA mutations in human disease. Nature Rev. Genet. 6, 389–402 (2005)

    Article  CAS  Google Scholar 

  13. Schaefer, A. M. et al. Prevalence of mitochondrial DNA disease in adults. Ann. Neurol. 63, 35–39 (2008)

    Article  CAS  Google Scholar 

  14. Steffann, J. et al. Analysis of mtDNA variant segregation during early human embryonic development: a tool for successful NARP preimplantation diagnosis. J. Med. Genet. 43, 244–247 (2006)

    Article  CAS  Google Scholar 

  15. Thorburn, D. R. & Dahl, H. H. Mitochondrial disorders: genetics, counseling, prenatal diagnosis and reproductive options. Am. J. Med. Genet. 106, 102–114 (2001)

    Article  CAS  Google Scholar 

  16. Byrne, J. A. et al. Producing primate embryonic stem cells by somatic cell nuclear transfer. Nature 450, 497–502 (2007)

    Article  ADS  CAS  Google Scholar 

  17. Mitalipov, S. M. et al. Reprogramming following somatic cell nuclear transfer in primates is dependent upon nuclear remodeling. Hum. Reprod. 22, 2232–2242 (2007)

    Article  CAS  Google Scholar 

  18. Wolf, D. P. et al. Use of assisted reproductive technologies in the propagation of rhesus macaque offspring. Biol. Reprod. 71, 486–493 (2004)

    Article  CAS  Google Scholar 

  19. Moraes, C. T., Atencio, D. P., Oca-Cossio, J. & Diaz, F. Techniques and pitfalls in the detection of pathogenic mitochondrial DNA mutations. J. Mol. Diagn. 5, 197–208 (2003)

    Article  CAS  Google Scholar 

  20. Tsuboniwa, N. et al. Safety evaluation of hemagglutinating virus of Japan—artificial viral envelope liposomes in nonhuman primates. Hum. Gene Ther. 12, 469–487 (2001)

    Article  CAS  Google Scholar 

  21. Poulton, J., Kennedy, S., Oakeshott, P. & Wells, D. Preventing transmission of maternally inherited mitochondrial DNA diseases. BMJ 338, b94 (2009)

    Article  Google Scholar 

  22. Roberts, R. M. Prevention of human mitochondrial (mtDNA) disease by nucleus transplantation into an enucleated donor oocyte. Am. J. Med. Genet. 87, 265–266 (1999)

    Article  CAS  Google Scholar 

  23. Cohen, J. et al. Ooplasmic transfer in mature human oocytes. Mol. Hum. Reprod. 4, 269–280 (1998)

    Article  CAS  Google Scholar 

  24. Lanzendorf, S. E. et al. Pregnancy following transfer of ooplasm from cryopreserved-thawed donor oocytes into recipient oocytes. Fertil. Steril. 71, 575–577 (1999)

    Article  CAS  Google Scholar 

  25. Cohen, J., Scott, R., Schimmel, T., Levron, J. & Willadsen, S. Birth of infant after transfer of anucleate donor oocyte cytoplasm into recipient eggs. Lancet 350, 186–187 (1997)

    Article  CAS  Google Scholar 

  26. Barritt, J. A., Brenner, C. A., Malter, H. E. & Cohen, J. Mitochondria in human offspring derived from ooplasmic transplantation. Hum. Reprod. 16, 513–516 (2001)

    Article  CAS  Google Scholar 

  27. Brenner, C. A., Barritt, J. A., Willadsen, S. & Cohen, J. Mitochondrial DNA heteroplasmy after human ooplasmic transplantation. Fertil. Steril. 74, 573–578 (2000)

    Article  CAS  Google Scholar 

  28. Brown, D. T. et al. Transmission of mitochondrial DNA disorders: possibilities for the future. Lancet 368, 87–89 (2006)

    Article  CAS  Google Scholar 

  29. Takeuchi, T., Neri, Q. V., Katagiri, Y., Rosenwaks, Z. & Palermo, G. D. Effect of treating induced mitochondrial damage on embryonic development and epigenesis. Biol. Reprod. 72, 584–592 (2005)

    Article  CAS  Google Scholar 

  30. Wilding, M. et al. Mitochondrial aggregation patterns and activity in human oocytes and preimplantation embryos. Hum. Reprod. 16, 909–917 (2001)

    Article  CAS  Google Scholar 

  31. Meirelles, F. V. & Smith, L. C. Mitochondrial genotype segregation in a mouse heteroplasmic lineage produced by embryonic karyoplast transplantation. Genetics 145, 445–451 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Meirelles, F. V. & Smith, L. C. Mitochondrial genotype segregation during preimplantation development in mouse heteroplasmic embryos. Genetics 148, 877–883 (1998)

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Sato, A. et al. Gene therapy for progeny of mito-mice carrying pathogenic mtDNA by nuclear transplantation. Proc. Natl Acad. Sci. USA 102, 16765–16770 (2005)

    Article  ADS  CAS  Google Scholar 

  34. Cree, L. M. et al. A reduction of mitochondrial DNA molecules during embryogenesis explains the rapid segregation of genotypes. Nature Genet. 40, 249–254 (2008)

    Article  CAS  Google Scholar 

  35. Wai, T., Teoli, D. & Shoubridge, E. A. The mitochondrial DNA genetic bottleneck results from replication of a subpopulation of genomes. Nature Genet. 40, 1484–1488 (2008)

    Article  CAS  Google Scholar 

Download references


The authors acknowledge the Division of Animal Resources, Surgical Team, Endocrine Technology Core, Imaging & Morphology Core and Molecular & Cellular Biology Core at the Oregon National Primate Research Center for providing expertise and services that contributed to this project. We are grateful to W. Sanger and M. Nelson for karyotyping services, C. Penedo for microsatellite analysis and S. Wong for providing Sendai virus. We thank J. Hennebold, R. Stouffer and D. Wolf for consulting, helpful discussions and critical reading of the manuscript. This study was supported by start-up funds from Oregon National Primate Research Center, Oregon Stem Cell Center and grants from the National Institutes of Health.

Author Contributions S.M. and M.T. conceived the study, designed experiments and conducted ST micromanipulations. M.S. performed ICSI, mitochondrial staining and analysis in oocytes. H.S., J.W. and Y.L. conducted ES cell derivation, characterization and differentiation. M.T., H.M., L.C., H.S. and Y.L. performed DNA/RNA isolations and mtDNA analyses. C.R. and O.K. conducted ovarian stimulations, oocyte recovery, ICSI and embryo transfers. S.M. and M.T. analysed the data and wrote the paper.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Shoukhrat Mitalipov.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-6 with Legends, Supplementary Tables 1-5, Supplementary Methods and Supplementary References. (PDF 1163 kb)

Supplementary Movie 1

This file shows the Mitochondrial Gene Replacement in Primate Offspring and Embryonic Stem Cells. (MOV 4490 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tachibana, M., Sparman, M., Sritanaudomchai, H. et al. Mitochondrial gene replacement in primate offspring and embryonic stem cells. Nature 461, 367–372 (2009).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


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.


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