Total oxidative status of mouse vitrified pre-antral follicles with pre-treatment of alpha lipoic acid.
Cryopreservation of pre-antral follicles is a hopeful technique to preserve female fertility. The aim of the present study was to evaluate reactive oxygen species (ROS) and total antioxidant capacity (TAC) levels of mouse vitrified pre-antral follicles in the presence of alpha lipoic acid (ALA).



Isolated pre-antral follicles (140-150 µm in diameter) were divided into vitrified-warmed and fresh groups. Each group was subjected to in vitro maturation with or without ALA for 12 days, followed by adding human chronic gonadotropin to induce ovulation. In vitro fertilization was performed to evaluate their developmental competence. In parallel, the amount of ROS and TAC were assessed after 0, 24, 48, 72, and 96 h of culture by 2',7'-dichlorofluorescin assay and ferric reducing/antioxidant power assay, respectively.



The respective rates of survival, antrum formation, and metaphase II oocytes were significantly higher in ALA-supplemented groups compared to the groups not treated with ALA. TAC and ROS levels were significantly decreased and increased, respectively during the culture period up to 96 h in the absence of ALA in both vitrified and non-vitrified samples. However, with pretreatment of ALA, TAC levels were increased significantly and remained constant up to 96 h in vitrified-warmed pre-antral follicles, while ROS levels completely returned to the level of starting point after 96 h of culture in the presence of ALA.



Pretreatment of ALA positively influences development of pre-antral follicles in vitrified and non-vitrified samples through increasing follicular TAC level and decreasing ROS levels.
School of Biology, Damghan University, Damghan, Iran. Zavareh.
Sahar
S
Hatami
School of Biology, Damghan University, Damghan, Iran. Zavareh.|Institute of Biological Sciences, Damghan University, Damghan, Iran.
Saeed
S
Zavareh
College of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Mojdeh
M
Salehnia
School of Biology, Damghan University, Damghan, Iran. Zavareh.|Institute of Biological Sciences, Damghan University, Damghan, Iran.
Taghi
T
Lashkarbolouki
School of Biology, Damghan University, Damghan, Iran. Zavareh.|Institute of Biological Sciences, Damghan University, Damghan, Iran.
Mohammad Taghi
MT
Ghorbanian
Laboratory of Molecular and Cellular Biology, Department of Basic Veterinary Sciences, School of Veterinary Medicine, Razi University, Kermanshah, Iran.
Isaac
I
Karimi
D000818:Animals; D000975:Antioxidants; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D051379:Mice; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D006080:Ovarian Follicle / Q000187:drug effects / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D008063:Thioctic Acid / Q000494:pharmacology*; D058989:Vitrification
D000975:Antioxidants; D017382:Reactive Oxygen Species; D008063:Thioctic Acid
Vitrification; Pre-antral follicle; Alpha lipoic acid; Reactive oxygen species; Total antioxidant capacity