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BMC Vet Res. 2019 Jan 17;15(1):31. doi: 10.1186/s12917-018-1743-2.

N-acetylcysteine addition after vitrification improves oocyte mitochondrial polarization status and the quality of embryos derived from vitrified murine oocytes.

Author information

1
Department of Cell Biology, School of Life Sciences, University of Extremadura, Avda. de Elvas, 6006, Badajoz, Spain.
2
Department of Physiology, Faculty of Nursing and Occupational Therapy, University of Extremadura, Avda. de la Universidad s/n, 10003, Cáceres, Spain.
3
Research Group of Intracellular Signaling and Technology of Reproduction (SINTREP), Institute of Biotechnology in Agriculture and Livestock (INBIO G+C) University of Extremadura, Avda. de la Universidad s/n, 10003, Cáceres, Spain.
4
Jesús Usón Minimally Invasive Surgery Centre, Carretera N-521, km. 41,8, 10071, Cáceres, Spain.
5
Jesús Usón Minimally Invasive Surgery Centre, Carretera N-521, km. 41,8, 10071, Cáceres, Spain. bea_macias@hotmail.com.
6
Research Group of Intracellular Signaling and Technology of Reproduction (SINTREP), Institute of Biotechnology in Agriculture and Livestock (INBIO G+C) University of Extremadura, Avda. de la Universidad s/n, 10003, Cáceres, Spain. bea_macias@hotmail.com.

Abstract

BACKGROUND:

Vitrification is the safest method to cryopreserve oocytes, however the process alters mitochondrial function resulting from increased reactive oxygen species (ROS) production. Our aim was to alleviate ROS stress in vitrified mice oocytes using N-acetylcysteine (NAC at 1 mM), to improve the oocyte's developmental competence.

RESULTS:

Hence, four experimental groups were compared: fresh oocytes (F-C), vitrified oocytes (V-C), NAC addition prior to oocyte vitrification (V-NAC-Pre) and NAC addition after vitrification (V-NAC-Post). V-NAC-Pre and V-NAC-Post exhibited higher levels of mitochondrial polarization compared to vitrified oocytes (36.5 ± 3.1, 37.7 ± 1.3 and 27.2 ± 2.4 measured as the spatial coefficient of variation/oocyte respectively, mean ± SEM; p < 0.05). However, ROS production increased in vitrified oocytes added with NAC compared to the vitrified control (1124.7 ± 102.1 [V-NAC-Pre] and 1063.2 ± 82.1 [V-NAC-Post] vs. 794.6 ± 164.9 [V-C]; arbitrary fluorescence units/oocyte, mean ± SEM; p < 0.05). ATP significantly decreased in V-NAC-Pre compared to V-NAC-Post oocytes (18.5 ± 6.9 vs. 54.2 ± 4.6 fmol/oocyte respectively, mean ± SEM; p < 0.05), and no differences were observed between V-NAC-Post, F-C and V-C groups. Blastocyst rates derived from F-C oocytes was higher than those derived from V-NAC-Pre (90.7 ± 1.8 vs. 79.1 ± 1.8, respectively, mean % ± SEM,; p < 0.05) but similar to those derived from V-NAC-Post (90.7 ± 1.8, mean % ± SEM, p > 0.05). Total blastomere count of blastocysts derived from V-NAC-Post after in vitro fertilization (IVF) was higher than embryos produced from V-C.

CONCLUSIONS:

The addition of NAC after vitrification improves the quality of vitrified mature murine oocytes while its addition prior to vitrification is advised against.

KEYWORDS:

Mouse; N-acetylcysteine; Oocyte; Vitrification

PMID:
30654800
PMCID:
PMC6337864
DOI:
10.1186/s12917-018-1743-2
[Indexed for MEDLINE]
Free PMC Article

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