Dmitry Nikiforov1,2, Cheng Junping3,4, Jesus Cadenas3, Vallari Shukla5, Robert Blanshard6,7, Susanne Elisabeth Pors3, Stine Gry Kristensen3, Kirsten Tryde Macklon8, Lotte Colmorn8, Erik Ernst9,10, Anne-Mette Bay-Bjørn9, Zeinab Ghezelayagh3, Yu Wakimoto3,11, Marie Louise Grøndahl12, Eva Hoffmann5, Claus Yding Andersen3. 1. Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children and Reproduction, Copenhagen University Hospital and Faculty of Health and Medical Sciences, University of Copenhagen, 2100, Copenhagen, Denmark. dmitry.nikiforov@regionh.dk. 2. Università degli studi di Teramo, 64100, Teramo, Italy. dmitry.nikiforov@regionh.dk. 3. Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children and Reproduction, Copenhagen University Hospital and Faculty of Health and Medical Sciences, University of Copenhagen, 2100, Copenhagen, Denmark. 4. People's Hospital of Guangxi Autonomous Region, Nanning, 530000, China. 5. DNRF Center for Chromosome Stability, Department of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, 2200, Copenhagen, Denmark. 6. Clinical Genomics Group, Illumina UK Ltd., Fulbourn, UK. 7. Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton, BN1 9RQ, UK. 8. The Fertility Clinic, Rigshospitalet, Copenhagen University Hospital, 2100, Copenhagen, Denmark. 9. The Fertility Clinic, Aarhus University Hospital, Skejby, 8200, Aarhus, Denmark. 10. The Fertility Clinic, Horsens Regional Hospital, 8700, Horsens, Denmark. 11. Department of Obstetrics and Gynecology, Hyogo College of Medicine, 663-8501 Nishi-nomiya, Hyogo, Japan. 12. The Fertility Clinic, Herlev Gentofte Hospital, 2740, Herlev, Denmark.
Abstract
PURPOSE: The aim of the present study was to improve the in vitro maturation (IVM) procedure using oocytes from surplus ovarian tissue after fertility preservation. METHODS: 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). RESULTS: 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. CONCLUSIONS: 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.
PURPOSE: The aim of the present study was to improve the in vitro maturation (IVM) procedure using oocytes from surplus ovarian tissue after fertility preservation. METHODS: 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 humanmidkine. Mature oocytes were assessed for aneuploidy using next-generation sequencing (NGS). RESULTS: 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. CONCLUSIONS: 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.
Entities:
Keywords:
Fertility preservation; Human immature oocytes; In vitro maturation; Oocyte diameter; Ovarian cryopreservation
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