Skip to main content
SpringerLink
Log in

Embryo development after mitochondrial supplementation from induced pluripotent stem cells

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The purpose of this study was to evaluate the effects of mitochondrial supplementation (MS) on early embryonic development and to assess the safety of MS treatments using induced pluripotent stem cells (iPSCs) as the mitochondrial donor.

Methods

In this study, we evaluated the effect of MS on early embryonic development using induced pluripotent stem cells (iPSCs) as the donor. Mouse zygotes were injected with either mitochondria from iPSCs or a vehicle solution. Several parameters were evaluated, including the rates of blastocyst formation and implantation, the weight of E13.5 embryos and placentas, the distribution of the donor mitochondrial DNA (mtDNA), and the pattern of methylation in the differentially methylated regions (DMRs) of the H19 and Snrpn genes.

Results

We found that neither the rates of blastocyst formation and implantation nor the weights of E13.5 embryos and placentas were significantly different between the MS and control groups. Additionally, the mtDNA from the iPSC donors could be detected in the muscle tissue of four fetuses and all placentas in the MS group. Finally, the methylation patterns of H19 and Snrpn DMRs remained unchanged by MS.

Conclusions

iPSC-derived mtDNA was directly involved in the process of embryonic development after MS. No adverse effects were seen when using iPSCs as a mitochondrial donor, but it remains to be seen whether this method can improve embryonic development, especially in older mice.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Dyall SD, Brown MT, Johnson PJ. Ancient invasions: from endosymbionts to organelles. Science. 2004;304:253–7.

    Article  CAS  PubMed  Google Scholar 

  2. de Bruin JP, Dorland M, Spek ER, Posthuma G, van Haaften M, Looman CW, Te VE. Age-related changes in the ultrastructure of the resting follicle pool in human ovaries. Biol Reprod. 2004;70:419–24.

    Article  PubMed  Google Scholar 

  3. Bentov Y, Casper RF. The aging oocyte—can mitochondrial function be improved? Fertil Steril. 2013;99:18–22.

    Article  CAS  PubMed  Google Scholar 

  4. St John J. Ooplasm donation in humans: the need to investigate the transmission of mitochondrial DNA following cytoplasmic transfer. Hum Reprod. 2002;17:1954–8.

  5. Seifer DB, DeJesus V, Hubbard K. Mitochondrial deletions in luteinized granulosa cells as a function of age in women undergoing in vitro fertilization. Fertil Steril. 2002;78:1046–8.

    Article  PubMed  Google Scholar 

  6. Perez GI, Trbovich AM, Gosden RG, Tilly JL. Mitochondria and the death of oocytes. Nature. 2000;403:500–1.

    Article  CAS  PubMed  Google Scholar 

  7. Yi YC, Chen MJ, Ho JY, Guu HF, Ho ES. Mitochondria transfer can enhance the murine embryo development. J Assist Reprod Genet. 2007;24:445–9.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Igarashi H, Takahashi T, Abe H, Nakano H, Nakajima O, Nagase S. Poor embryo development in post-ovulatoryin vivo-aged mouse oocytes is associated with mitochondrial dysfunction, but mitochondrial transfer from somatic cells is not sufficient for rejuvenation. Hum Reprod. 2016;31:2331–8.

    Article  PubMed  Google Scholar 

  9. Oktay K, Baltaci V, Sonmezer M, Turan V, Unsal E, Baltaci A, et al. Oogonial precursor cell-derived autologous mitochondria injection to improve outcomes in women with multiple IVF failures due to low oocyte quality: a clinical translation. Reprod Sci. 2015;22:1612–7.

    Article  PubMed  Google Scholar 

  10. Leighton PA, Ingram RS, Eggenschwiler J, Efstratiadis A, Tilghman SM. Disruption of imprinting caused by deletion of the H19 gene region in mice. Nature. 1995;375:34–9.

    Article  CAS  PubMed  Google Scholar 

  11. Abu-Amero S, Monk D, Frost J, Preece M, Stanier P, Moore GE. The genetic aetiology of Silver-Russell syndrome. J Med Genet. 2008;45:193–9.

    Article  CAS  PubMed  Google Scholar 

  12. Leff SE, Brannan CI, Reed ML, Ozcelik T, Francke U, Copeland NG, Jenkins NA. Maternal imprinting of the mouse Snrpn gene and conserved linkage homology with the human Prader-Willi syndrome region. Nat Genet. 1992;2:259–64.

    Article  CAS  PubMed  Google Scholar 

  13. Han J, Yuan P, Yang H, Zhang J, Soh BS, Li P, et al. Tbx3 improves the germ-line competency of induced pluripotent stem cells. Nature. 2010;463:1096–100.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Hashizume O, Ohnishi S, Mito T, Shimizu A, Iashikawa K, Nakada K, et al. Epigenetic regulation of the nuclear-coded GCAT and SHMT2 genes confers human age-associated mitochondrial respiration defects. Sci Rep-UK. 2015;5:10434.

    Article  Google Scholar 

  15. Ma H, Folmes CD, Wu J, Morey R, Mora-Castilla S, Ocampo A, et al. Metabolic rescue in pluripotent cells from patients with mtDNA disease. Nature. 2015;524(7564):234–8.

    Article  CAS  PubMed  Google Scholar 

  16. Hasuwa H, Muro Y, Ikawa M, Kato N, Tsujimoto Y, Okabe M. Transgenic mouse sperm that have green acrosome and red mitochondria allow visualization of sperm and their acrosome reaction in vivo. Exp Anim. 2010;59:105–7.

    Article  CAS  PubMed  Google Scholar 

  17. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–76.

    Article  CAS  PubMed  Google Scholar 

  18. Pei Y, Yue L, Zhang W, Wang Y, Wen B, Zhong L, et al. Improvement in mouse iPSC induction by Rab32 reveals the importance of lipid metabolism during reprogramming. Sci Rep. 2015;5:16539.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Takeda K. Microinjection of cytoplasm or mitochondria derived from somatic cells affects parthenogenetic development of murine oocytes. Biol Reprod. 2005;72:1397–404.

    Article  CAS  PubMed  Google Scholar 

  20. Nagai S, Kasai T, Hirata S, Hoshi K, Yanagimachi R, Huang T. Cytoplasmic transfer in the mouse in conjunction with intracytoplasmic sperm injection. Reprod BioMed Online. 2004;8:75–80.

    Article  PubMed  Google Scholar 

  21. Yi Y, Chen M, Ho JY, Guu H, Ho ES. Mitochondria transfer can enhance the murine embryo development. J Assist Reprod Gen. 2007;24:445–9.

    Article  Google Scholar 

  22. Luo SM, Ge ZJ, Wang ZW, Jiang ZZ, Wang ZB, Ouyang YC, et al. Unique insights into maternal mitochondrial inheritance in mice. Proc Natl Acad Sci. 2013;110:13038–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Cheng K, Fu X, Zhang R, Jia G, Hou Y, Zhu S. Effect of oocyte vitrification on deoxyribonucleic acid methylation of H19, Peg3, and Snrpn differentially methylated regions in mouse blastocysts. Fertil Steril. 2014;102:1183–90.

    Article  CAS  PubMed  Google Scholar 

  24. Hao Y, Wax D, Zhong Z, Murphy C, Ross JW, Rieke A, et al. Porcine skin-derived stem cells can serve as donor cells for nuclear transfer. Cloning Stem Cells. 2009;11:101–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Reddy P, Ocampo A, Suzuki K, Luo J, Bacman SR, Williams SL, et al. Selective elimination of mitochondrial mutations in the germline by genome editing. Cell. 2015;161:459–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Shimizu A, Mito T, Hayashi C, Ogasawara E, Koba R, Negishi I, et al. Transmitochondrial mice as models for primary prevention of diseases caused by mutation in the tRNA(Lys) gene. Proc Natl Acad Sci U S A. 2014;111:3104–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Yang L, Long Q, Liu J, Tang H, Li Y, Bao F, et al. Mitochondrial fusion provides an ‘initial metabolic complementation’ controlled by mtDNA. Cell Mol Life Sci. 2015;72:2585–98.

    Article  CAS  PubMed  Google Scholar 

  28. Motta PM, Nottola SA, Makabe S, Heyn R. Mitochondrial morphology in human fetal and adult female germ cells. Hum Reprod. 2000;15(Suppl 2):129–47.

    Article  PubMed  Google Scholar 

  29. Cagnone GL, Tsai TS, Makanji Y, Matthews P, Gould J, Bonkowski MS, et al. Restoration of normal embryogenesis by mitochondrial supplementation in pig oocytes exhibiting mitochondrial DNA deficiency. Sci Rep. 2016;6:23229.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Morrow EH, Reinhardt K, Wolff JN, Dowling DK. Risks inherent to mitochondrial replacement. EMBO Rep. 2015;16:541–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Lee HS, Ma H, Juanes RC, Tachibana M, Sparman M, Woodward J, et al. Rapid mitochondrial DNA segregation in primate preimplantation embryos precedes somatic and germline bottleneck. Cell Rep. 2012;1:506–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant Nos. 81370680, 81402168, and 31571497), the Specialized Research Fund for the Doctoral Program of the Chinese Ministry of Education (Grant No. 20130171130009), and the Natural Science Foundation’s Key Research Project of Guangdong Province (Grant No. 2013020012660).

Author information

Author notes

  1. Ruiqi Li and Bingqiang Wen contributed equally to this work.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Obstetrics and Gynecology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120, People’s Republic of China

    Ruiqi Li, Haijing Zhao, Nengyong Ouyang, Songbang Ou, Wenjun Wang & Dongzi Yang

  2. State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, People’s Republic of China

    Bingqiang Wen & Jianyong Han

  3. Reproductive Medicine Centre, Department of Obstetrics and Gynaecology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120, People’s Republic of China

    Dongzi Yang

Authors
  1. Ruiqi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bingqiang Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Haijing Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nengyong Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Songbang Ou
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wenjun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jianyong Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dongzi Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Ruiqi Li analyzed the data and drafted the manuscript. Bingqiang Wen induced iPSCs and evaluated the pluripotency of iPSCs. Haijing Zhao also drafted the manuscript. Nengyong Ouyang, Songbang O, and Meiqi Mai collected the data. Wenjun Wang revised the manuscript. Jianyong Han and Dongzi Yang conceived and designed the study. All authors interpreted the data.

Corresponding authors

Correspondence to Jianyong Han or Dongzi Yang.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethics approval

All animal protocols were approved by and performed in accordance with the requirements of the Institutional Animal Care and Use Committee at the authors’ university.

Electronic supplementary material

Supplementary Figure 1

The pluripotency detection of iPSCs. A) AP test; B) three germ layersof the teratoma; C) immunostaining of OCT4 and SOX2. (JPEG 8614 kb)

Supplementary Figure 2

Mitochondria ultrastructure and morphology of iPSCs. IPSCs mitochondria were immature, with round or oval shapes, possess few cristae and have an electron dense matrix. Mitochondria (arrow), nucleus (*) (12000×).. (JPEG 15112 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, R., Wen, B., Zhao, H. et al. Embryo development after mitochondrial supplementation from induced pluripotent stem cells. J Assist Reprod Genet 34, 1027–1033 (2017). https://doi.org/10.1007/s10815-017-0948-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-017-0948-9

Keywords

Search

Navigation