Past Issue

Volume 9, Number 4, Jan-Mar 2016, Pages: 442-451

Effect of In Vitro Maturation Technique and Alpha Lipoic Acid Supplementation on Oocyte Maturation Rate: Focus on Oxidative Status of Oocytes


Saeed Zavareh, Ph.D, 1, 2, *, Isaac Karimi, D.V.M., D.V.Sc, 3, Mojdeh Salehnia, Ph.D, 4, Ali Rahnama, M.Sc, 1,
School of Biology, Damghan University, Damghan, Iran
Institute of Biological Sciences, Damghan University, Damghan, Iran
Laboratory of Molecular and Cellular Biology, Department of Basic Veterinary Sciences, School of Veterinary Medicine, Razi University, Kermanshah, Iran
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
* Corresponding Address: P.O. Box: 41167-36719 School of Biology Damghan University Ceshme Ali Street Damghan Iran Email:Zavareh.S@du.ac.ir

Abstract

Background

Therapeutic potential of in vitro maturation (IVM) in infertility is growing with great promise. Although significant progress is obtained in recent years, existing IVM protocols are far from favorable results. The first aim of this study was to investigate whether two step IVM manner change reactive oxygen species (ROS) and total anti- oxidant capacity (TAC) levels. The second aim was to find the effect of alpha lipoic acid (ALA) supplementation on oocyte maturation rate and on ROS/TAC levels during IVM.

Materials and Methods

In this experimental study, mouse germinal vesicle (GV) oocytes divided into cumulus denuded oocytes (DOs) and cumulus oocyte complexes (COCs) groups. GVs were matured in vitro in the presence or absence of ALA only for 18 hours (control) or with pre-culture of forskolin plus cilostamide for an additional 18 hours. Matured oocytes obtained following 18 and 36 hours based on experimental design. In parallel, the ROS and TAC levels were measured at different time (0, 18 and 36 hours) by 2',7'-dichlorodihydrofluorescein (DCFH) probe and ferric reducing/antioxidant power (FRAP) assay, respectively.

Results

Maturation rate of COCs was significantly higher than DOs in control group (P<0.05), while there was no significant difference between COCs and DOs when were pre-cultured with forskolin plus cilostamide. ROS and TAC levels was increased and decreased respectively in DOs after 18 hours while in COCs did not change at 18 hours and showed a significant increase and decrease respectively at 36 hours (P<0.05). ROS and TAC levels in the presence of ALA were significantly decreased and increased respectively after 36 hours (P<0.05) whereas, maturation rates of COCs and DOs were similar to their corresponding control groups.

Conclusion

ALA decreased ROS and increased TAC but could not affect maturation rate of both COCs and DOs in one or two step IVM manner.