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Nuclear genome transfer in human oocytes eliminates mitochondrial DNA variants

Nature volume 493, pages 632–637 (2013)Cite this article

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Abstract

Mitochondrial DNA mutations transmitted maternally within the oocyte cytoplasm often cause life-threatening disorders. Here we explore the use of nuclear genome transfer between unfertilized oocytes of two donors to prevent the transmission of mitochondrial mutations. Nuclear genome transfer did not reduce developmental efficiency to the blastocyst stage, and genome integrity was maintained provided that spontaneous oocyte activation was avoided through the transfer of incompletely assembled spindle–chromosome complexes. Mitochondrial DNA transferred with the nuclear genome was initially detected at levels below 1%, decreasing in blastocysts and stem-cell lines to undetectable levels, and remained undetectable after passaging for more than one year, clonal expansion, differentiation into neurons, cardiomyocytes or β-cells, and after cellular reprogramming. Stem cells and differentiated cells had mitochondrial respiratory chain enzyme activities and oxygen consumption rates indistinguishable from controls. These results demonstrate the potential of nuclear genome transfer to prevent the transmission of mitochondrial disorders in humans.

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Figure 1: Efficient development and genomic integrity after nuclear genome exchange.
Figure 2: Spontaneous activation can be prevented through spindle cooling.
Figure 3: Low levels of mtDNA carryover.
Figure 4: swaPS cells support a normal metabolic profile.

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Gene Expression Omnibus

Data deposits

Illumina array data have been deposited at the Gene Expression Omnibus (GEO) under accession number GSE42077; Affymetrix array data have been deposited at the GEO under accession number GSE42271.

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Acknowledgements

We thank S. Chang and K. Eggan for discussions, Z. Hall for critical reading of the manuscript, and L. Yu and O. Nahum for SNP-array preparation. We thank anonymous oocyte donors for participating in research, and M. Spencer for a Lykos laser system. This work was supported by the New York Stem Cell Foundation, the New York State Stem Cell Science award C026184, and the Bernard and Anne Spitzer Fund.

Author information

Author notes

  1. Mark V. Sauer and Dieter Egli: These authors contributed equally to this work.

Authors and Affiliations

  1. The New York Stem Cell Foundation Laboratory, New York, 10032, USA

    Daniel Paull, Keren A. Weiss, Latoya Stewart, Haiqing Hua, Matthew Zimmer, David J. Kahler, Scott A. Noggle & Dieter Egli

  2. Department of Neurology, Columbia University, New York, 10032, USA

    Valentina Emmanuele & Michio Hirano

  3. Reproductive Medicine Associates of New Jersey, New Jersey, 07960, USA

    Nathan Treff

  4. Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, 10032, USA

    Haiqing Hua & Robin S. Goland

  5. Center for Women’s Reproductive Care, College of Physicians and Surgeons, Columbia University, New York, 10019, USA

    Robert Prosser & Mark V. Sauer

  6. Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York, 10032, USA

    Mark V. Sauer

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Contributions

M.V.S. consented oocyte donors and retrieved oocytes. R.P. contributed IVF developmental data. R.S.G. and M.V.S. wrote institutional review board and consent documents. D.E., D.P. and S.N. designed and performed experiments with oocytes. D.P. and V.E. determined heteroplasmy. N.T. performed array analysis of single cells. D.E., D.P., V.E., L.S., K.A.W., H.H., M.Z. and D.J.K characterized stem-cell lines. D.E., D.P., V.E. and M.H. wrote the paper.

Corresponding authors

Correspondence to Mark V. Sauer or Dieter Egli.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-9, Supplementary Tables 1-8 and Supplementary Karyotypes. (PDF 41490 kb)

SwaPS1 cardiomyocytes

This video shows a contracting embryoid body that had undergone directed differentiation toward a cardiac (mesodermal) lineage. (MOV 949 kb)

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Paull, D., Emmanuele, V., Weiss, K. et al. Nuclear genome transfer in human oocytes eliminates mitochondrial DNA variants. Nature 493, 632–637 (2013). https://doi.org/10.1038/nature11800

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