Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 12, pp 1587–1594 | Cite as

Mitochondrial DNA content is associated with ploidy status, maternal age, and oocyte maturation methods in mouse blastocysts

  • Xin TaoEmail author
  • Jessica N. Landis
  • Rebecca L. Krisher
  • Francesca E. Duncan
  • Elena Silva
  • Agnieszka Lonczak
  • Richard T. ScottIII
  • Yiping Zhan
  • Tinchun Chu
  • Richard T. ScottJr
  • Nathan R. Treff
Embryo Biology



It was reported that mitochondrial DNA (mtDNA) was significantly increased in aneuploid human embryos compared to euploid embryos and was also associated with maternal age. In this study, we further established the mouse model of mtDNA quantitation in reproductive samples based on whole-genome amplification (WGA) and next-generation sequencing (NGS).


WGA followed by NGS-based mtDNA quantitation was first performed on 6 single- and 100-cell samples from a tumor-derived mouse cell line, which was exposed to ethidium bromide to reduce mtDNA content. The relative mtDNA content was normalized to nuclear DNA. This method was then applied to mouse reproductive samples, including 40 pairs of oocytes and polar bodies from 8 CD-1 female mice of advanced reproductive age and 171 blastocysts derived via in vitro maturation (IVM) or in vivo maturation (IVO) from young (6–9 weeks) and reproductively aged (13.5 months) female CF-1 mice.


Exposure to ethidium bromide for 3 and 6 days decreased mtDNA levels in both the single- and 100-cell samples as expected. Results demonstrated that the first polar body contained an average of 0.9% of mtDNA relative to oocytes. Compared to the cells in blastocysts, oocytes contained about 180 times as much mtDNA per cell. mtDNA levels were compared among blastocysts from reproductively young and old female mice that had either been produced by IVM or IVO. Cells in blastocysts from younger mice contained significantly lower amounts of mtDNA compared to aged mice (P < 0.0001). Cells in blastocysts produced via IVO had higher mtDNA content than IVM-derived blastocysts (P = 0.0001). Cells in aneuploid blastocysts were found to have significantly higher (1.74-fold) levels of mtDNA compared to euploid blastocysts (P = 0.0006).


A reliable method for assessing mtDNA content in mouse gametes and embryos was established. Relative mtDNA levels were elevated in aneuploid embryos relative to euploid embryos, were higher in blastocysts from reproductively old mice relative to young mice, and were lower in embryos derived from IVM compared to IVO.


Aging Aneuploidy Mouse blastocyst Mitochondrial DNA NGS 


Compliance with ethical standards

All animal experiments were approved by the Institutional Animal Care and Use Committee (Northwestern University) or Colorado Center for Reproductive Medicine Ethics in Research Committee and were carried out in accordance with National Institutes of Health Guidelines and the Society for the Study of Reproduction’s specific guidelines and standards.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Xin Tao
    • 1
    • 2
    Email author
  • Jessica N. Landis
    • 1
  • Rebecca L. Krisher
    • 3
  • Francesca E. Duncan
    • 4
  • Elena Silva
    • 3
  • Agnieszka Lonczak
    • 1
  • Richard T. ScottIII
    • 1
  • Yiping Zhan
    • 1
  • Tinchun Chu
    • 2
  • Richard T. ScottJr
    • 5
  • Nathan R. Treff
    • 6
  1. 1.The Foundation for Embryonic CompetenceBasking RidgeUSA
  2. 2.Department of Biological SciencesSeton Hall UniversitySouth OrangeUSA
  3. 3.Colorado Center for Reproductive MedicineLone TreeUSA
  4. 4.Northwestern UniversityChicagoUSA
  5. 5.Reproductive Medicine Associates of New JerseyBasking RidgeUSA
  6. 6.Rutgers, the State University of New JerseyNew BrunswickUSA

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