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Improving the maturation rate of human oocytes collected ex vivo during the cryopreservation of ovarian tissue



The aim of the present study was to improve the in vitro maturation (IVM) procedure using oocytes from surplus ovarian tissue after fertility preservation.


Twenty-five patients aged 17–37 years were included in the study. Maturation was compared between oocytes collected in HEPES-buffered medium or saline, and we determined whether transport on ice prior to oocyte collection affected maturation. Two different IVM media were used that were supplemented with and without recombinant human midkine. Mature oocytes were assessed for aneuploidy using next-generation sequencing (NGS).


On average, 36 immature oocytes were collected from each patient (range 7–90, N = 895). Oocytes recovered from HEPES-buffered medium matured at a higher rate than oocytes recovered from saline (36% vs 26%, p < 0.01). Ovarian transportation on ice prior to the procedure negatively affected maturation compared with non-transported samples (42% vs 27%, p < 0.01). The addition of midkine improved maturation rate (34% vs 27%, p < 0.05). On average, 11 MII oocytes were obtained per patient (range 1–30). NGS of 53 MII oocytes and their first polar bodies indicated that 64% were euploid.


The study demonstrated unexpectedly high number of immature oocytes collected from surplus ovarian tissue without any stimulation. The overall MII rate was one in three, resulting in a total number of MII oocytes that was similar to the number obtained after ovarian stimulation. If these MII oocytes prove suitable for IVF, they will provide a substantial improvement in fertility preservation for patients and advance IVM as an interesting platform for further improvements in assisted reproduction.

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The grants from the organizations listed below are gratefully acknowledged as sources of financial support. ERH, VS and RB are grateful for the support from The Independent Research Fund Denmark, the NNF Young Investigator Award (grant number 16662), the ERC (grant number 724718) and the Danish National Research Foundation (DNRF115, 6110-00344B). The project ITN REP-BIOTECH 675526 funded by the European Union, European Joint Doctorate in Biology and Technology of the Reproductive Health is acknowledged for supporting DN. The University Hospital of Copenhagen, the EU Interregional Project ReproUnion and Vera and Carl Johan Michaelsens Legat are acknowledged for providing support for CYA, SGK and SEP. All personnel, including the staff involved in clinical activities in fertility preservation, are acknowledged for their passionate work.

Author information




CYA, DN, CJ, SEP and SGK planned the study. DN, CJ, VS, RB, SEP, SGK, MLG performed the experiments and collected the data. DN, CJ, SEP, SGK, JC, VS, RB, ZG, YW and EH analysed the data and produced figures. KTM, LC, EE and AMBB recruited patients and collected the tissues. DN, SEP, SGK and CYA wrote the manuscript. All authors approved the manuscript.

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Correspondence to Dmitry Nikiforov.

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Nikiforov, D., Junping, C., Cadenas, J. et al. Improving the maturation rate of human oocytes collected ex vivo during the cryopreservation of ovarian tissue. J Assist Reprod Genet 37, 891–904 (2020).

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  • Human immature oocytes
  • Fertility preservation
  • Ovarian cryopreservation
  • In vitro maturation
  • Oocyte diameter