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.

  • Article
  • Published:

Towards germline gene therapy of inherited mitochondrial diseases


Mutations in mitochondrial DNA (mtDNA) are associated with severe human diseases and are maternally inherited through the egg’s cytoplasm. Here we investigated the feasibility of mtDNA replacement in human oocytes by spindle transfer (ST; also called spindle–chromosomal complex transfer). Of 106 human oocytes donated for research, 65 were subjected to reciprocal ST and 33 served as controls. Fertilization rate in ST oocytes (73%) was similar to controls (75%); however, a significant portion of ST zygotes (52%) showed abnormal fertilization as determined by an irregular number of pronuclei. Among normally fertilized ST zygotes, blastocyst development (62%) and embryonic stem cell isolation (38%) rates were comparable to controls. All embryonic stem cell lines derived from ST zygotes had normal euploid karyotypes and contained exclusively donor mtDNA. The mtDNA can be efficiently replaced in human oocytes. Although some ST oocytes displayed abnormal fertilization, remaining embryos were capable of developing to blastocysts and producing embryonic stem cells similar to controls.

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

Access options

Buy this article

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

Figure 1: Experimental design and main outcomes after ST with human oocytes.
Figure 2: Abnormal pronuclear formation and spindle morphology in human ST zygotes.
Figure 3: Genetic analysis of ESCs derived from human ST embryos.

Similar content being viewed by others


  1. Gropman, A. L. Diagnosis and treatment of childhood mitochondrial diseases. Curr. Neurol. Neurosci. Rep. 1, 185–194 (2001)

    Article  CAS  Google Scholar 

  2. Haas, R. H. et al. Mitochondrial disease: a practical approach for primary care physicians. Pediatrics 120, 1326–1333 (2007)

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Elliott, H. R., Samuels, D. C., Eden, J. A., Relton, C. L. & Chinnery, P. F. Pathogenic mitochondrial DNA mutations are common in the general population. Am. J. Hum. Genet. 83, 254–260 (2008)

    Article  CAS  Google Scholar 

  5. Tachibana, M., Sparman, M. & Mitalipov, S. Chromosome transfer in mature oocytes. Nature Protocols 5, 1138–1147 (2010)

    Article  CAS  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  7. 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 

  8. Danan, C. et al. Evaluation of parental mitochondrial inheritance in neonates born after intracytoplasmic sperm injection. Am. J. Hum. Genet. 65, 463–473 (1999)

    Article  CAS  Google Scholar 

  9. Lee, H.-S. et al. Rapid mitochondrial DNA segregation in primate preimplantation embryos precedes somatic and germline bottleneck. Cell Reports 1, 506–515 (2012)

    Article  CAS  Google Scholar 

  10. Burgstaller, J. P., Schinogl, P., Dinnyes, A., Muller, M. & Steinborn, R. Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer. BMC Dev. Biol. 7, 141 (2007)

    Article  Google Scholar 

  11. Wakayama, T. & Yanagimachi, R. The first polar body can be used for the production of normal offspring in mice. Biol. Reprod. 59, 100–104 (1998)

    Article  CAS  Google Scholar 

  12. Susko-Parrish, J. L., Leibfried-Rutledge, M. L., Northey, D. L., Schutzkus, V. & First, N. L. Inhibition of protein kinases after an induced calcium transient causes transition of bovine oocytes to embryonic cycles without meiotic completion. Dev. Biol. 166, 729–739 (1994)

    Article  CAS  Google Scholar 

  13. Forman, E. J. et al. Oocyte vitrification does not increase the risk of embryonic aneuploidy or diminish the implantation potential of blastocysts created after intracytoplasmic sperm injection: a novel, paired randomized controlled trial using DNA fingerprinting. Fertil. Steril. 98, 644–649 (2012)

    Article  CAS  Google Scholar 

  14. Rienzi, L. et al. Consistent and predictable delivery rates after oocyte vitrification: an observational longitudinal cohort multicentric study. Hum. Reprod. 27, 1606–1612 (2012)

    Article  Google Scholar 

  15. Moreno-Loshuertos, R. et al. Differences in reactive oxygen species production explain the phenotypes associated with common mouse mitochondrial DNA variants. Nature Genet. 38, 1261–1268 (2006)

    Article  CAS  Google Scholar 

  16. Fisher, D. L., Brassac, T., Galas, S. & Doree, M. Dissociation of MAP kinase activation and MPF activation in hormone-stimulated maturation of Xenopus oocytes. Development 126, 4537–4546 (1999)

    CAS  PubMed  Google Scholar 

  17. Runft, L. L., Jaffe, L. A. & Mehlmann, L. M. Egg activation at fertilization: where it all begins. Dev. Biol. 245, 237–254 (2002)

    Article  CAS  Google Scholar 

  18. Mitalipov, S. M., Nusser, K. D. & Wolf, D. P. Parthenogenetic activation of rhesus monkey oocytes and reconstructed embryos. Biol. Reprod. 65, 253–259 (2001)

    Article  CAS  Google Scholar 

  19. Gao, S., Han, Z., Kihara, M., Adashi, E. & Latham, K. E. Protease inhibitor MG132 in cloning: no end to the nightmare. Trends Biotechnol. 23, 66–68 (2005)

    Article  CAS  Google Scholar 

  20. Kikuchi, K. et al. Maturation/M-phase promoting factor: a regulator of aging in porcine oocytes. Biol. Reprod. 63, 715–722 (2000)

    Article  CAS  Google Scholar 

  21. Mandelbaum, J. et al. Effects of cryopreservation on the meiotic spindle of human oocytes. Eur. J. Obstet. Gynecol. Reprod. Biol. 113 (Suppl 1). S17–S23 (2004)

    Article  Google Scholar 

  22. Smith, D. G. Genetic characterization of Indian-origin and Chinese-origin rhesus macaques (Macaca mulatta). Comp. Med. 55, 227–230 (2005)

    CAS  PubMed  Google Scholar 

  23. Donnez, J. et al. Children born after autotransplantation of cryopreserved ovarian tissue. a review of 13 live births. Ann. Med. 43, 437–450 (2011)

    Article  Google Scholar 

  24. Lee, D. M. et al. Live birth after ovarian tissue transplant. Nature 428, 137–138 (2004)

    Article  ADS  CAS  Google Scholar 

Download references


The authors would like to acknowledge the Oregon Health & Science University (OHSU) Embryonic Stem Cell Research Oversight Committee and the Institutional Review Board for providing oversight and guidance. We thank oocyte and sperm donors and staff at the Women’s Health Research Unit at the Center for Women’s Health, University Fertility Consultants and Reproductive Endocrinology & Infertility Division at the Department of Obstetrics & Gynecology of Oregon Health & Science University for their support and procurement of human gametes. The Division of Animal Resources, Surgery Team, Assisted Reproductive Technology & Embryonic Stem Cell Core, Endocrine Technology Core, Imaging & Morphology Core, Flow Cytometry Core and Molecular & Cellular Biology Core at the Oregon National Primate Research Center provided expertise and services for the nonhuman primate research. Hamilton Thorne Inc., donated an XYClone laser system for this study. We are grateful to W. Sanger and D. Zaleski for karyotyping services, C. Penedo for microsatellite analysis and J. Hennebold for consulting on metabolic assays. We are also indebted to A. Steele, R. Cervera Juanes and E. Wolff for their technical support. The human oocyte/embryo research was supported by grants from the OHSU Center for Women’s Health Circle of Giving and other OHSU institutional funds, as well as the Leducq Foundation. The nonhuman primate study was supported by grants from the National Institutes of Health HD063276, HD057121, HD059946, EY021214 and 8P51OD011092.

Author information

Authors and Affiliations



M.T., P.A., J.J. and S.M. conceived the study, designed experiments and wrote institutional review board protocols. P.A., M.S. and N.M.G. coordinated recruitment of participants. P.A., K.M., D.B., D.L., D.W. and P.P. performed ovarian stimulation and oocyte recovery. M.T. conducted ST micromanipulations. M.S., K.M. and S.M. performed ICSI. M.T., M.S., J.W., D.M.S., N.M.G., R.T.-H. and E.K. conducted ESC derivation and characterization. S.G. analysed teratoma tumours. M.T., H.M. and D.M.S. performed DNA/RNA isolations, metabolic and mtDNA analyses. C.R., M.T., M.S.,H.-S.L., R.S. and S.M. conducted monkey studies. M.T., R.S., J.J., P.P. and S.M. analysed data and wrote the paper.

Corresponding author

Correspondence to Shoukhrat Mitalipov.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-10, Supplementary Tables 1-7, Supplementary Methods and Supplementary References. (PDF 1329 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tachibana, M., Amato, P., Sparman, M. et al. Towards germline gene therapy of inherited mitochondrial diseases. Nature 493, 627–631 (2013).

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: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research