Equilibration time with cryoprotectants, but not melatonin supplementation during in vitro maturation, affects viability and metaphase plate morphology of vitrified porcine mature oocytes.

Autor: Gonzalez-Plaza A; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Brullo C; Department of Veterinary Medical Sciences and CIRI-SDV, University of Bologna, Bologna, Italy., Cambra JM; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Garcia M; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Iacono E; Department of Veterinary Medical Sciences and CIRI-SDV, University of Bologna, Bologna, Italy., Parrilla I; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Gil MA; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Martinez EA; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain., Martinez CA; Department of Biomedical & Clinical Sciences (BKV), BKH/Obstetrics & Gynaecology, Faculty of Medicine and Health Sciences, Linköping University, Linköping, Sweden., Cuello C; Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, Murcia, Spain.; Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Murcia, Spain.
Jazyk: angličtina
Zdroj: Reproduction in domestic animals = Zuchthygiene [Reprod Domest Anim] 2022 Oct; Vol. 57 Suppl 5, pp. 58-63. Date of Electronic Publication: 2022 May 24.
DOI: 10.1111/rda.14158
Abstrakt: The aims of this study were to investigate the effects of different equilibration times with cryoprotectants on viability and metaphase plate morphology of vitrified-warmed porcine mature oocytes (Experiment 1) and to evaluate the effects of supplementation with 10 -9 M melatonin during in vitro maturation on these parameters (Experiment 2). In Experiment 1, 2,392 mature oocytes were vitrified using different equilibration times of oocytes with cryoprotectants (3, 10, 15, 20, 30, 40, 60 and 80 min). Fresh oocytes matured in vitro for 44 hr (n = 509) were used as controls. In Experiment 2, a total of 573 COCs were used. COCs were matured with 10 -9 M melatonin supplementation or without melatonin (control). Some oocytes from each group were vitrified with a 60-min equilibration time with cryoprotectants according to the results of Experiment 1. The remaining oocytes from each maturation group were used as fresh control groups. In both experiments, oocytes were stained with 2',7'-dichlorodihydrofuorescein diacetate and Hoechst 33342 to assess viability and metaphase plate morphology, respectively. Vitrification and warming affected (p < .01) oocyte viability compared with controls, which were all viable after 44 hr of IVM. In Experiment 1, the longer the equilibration time with cryoprotectants, the higher the viability. Oocytes equilibrated for 60 and 80 min had the highest (p < .05) viability and similar metaphase plate characteristics to the fresh control oocytes. In Experiment 2, supplementation with melatonin during in vitro maturation had no effect on oocyte viability or metaphase plate morphology of vitrified-warmed oocytes. In conclusion, under our experimental conditions, vitrified porcine mature oocytes equilibrated with cryoprotectants for 60 or 80 min exhibited the highest viability and similar metaphase plate characteristics to fresh controls. Furthermore, supplementation with 10 -9 M melatonin during in vitro maturation had no effect on these parameters.
(© 2022 The Authors. Reproduction in Domestic Animals published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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