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. 2017 Feb;11(2):BC10-BC14.
doi: 10.7860/JCDR/2017/21778.9298. Epub 2017 Feb 1.

Effect of Antioxidants (β-mercaptoethanol and Cysteamine) on Assisted Reproductive Technology In vitro

Mohsen Nikseresht  1 , Mehdi Akbartabar Toori  1 , Hamid Reza Rahimi  2 , Ali Reza Fallahzadeh  3 , Iraj Ragerdi Kahshani  4 , Seyedeh Fatemeh Hashemi  5 , Solmaz Bahrami  5 , Reza Mahmoudi  6
Affiliations

Effect of Antioxidants (β-mercaptoethanol and Cysteamine) on Assisted Reproductive Technology In vitro

Mohsen Nikseresht et al. J Clin Diagn Res. 2017 Feb.

Abstract

Introduction: Oocyte Culture of Germinal Vesicle (GV) and its growth improves Assisted Reproductive Technology (ART) invitro and infertility. Inappropriate culture medium environment, low quality of oocytes, increase in Oxidative Stress (OS) events, Reactive Oxygen Species (ROS) and free radicals production are the main factors that result in unsuccessful Invitro Maturation (IVM) and decrease in reproduction.

Aim: The present study was conducted with the aim to evaluate the effect of β-mercaptoethanol (BME) and Cysteamine (CYS) on IVM improvement, embryo fertilization and development of blastocyst of mouse immature oocyte.

Materials and methods: Oocytes were obtained from 4-6 weeks old Naval Medical Research Institute (NMRI) female mice, 48 hours after stimulation with Intraperitoneal (IP) injection of 10 IU Pregnant Mare Serum Gonadotropin (PMSG). GV oocyte with and without cumulus cells were isolated from ovaries and cultured in Tissue Culture Medium (TCM) 199 with availability of 100 μM of antioxidants (BME and CYS). After 24 hours, mature oocyte in metaphase II (MII) were fertilized with sperm in In vitro Fertilization (IVF) medium (T6) and evaluated for fetal development into blastocyst.

Results: BME and CYS could significantly (p<0.05) increase the rate of IVM and oocyte evolution, and embryo formation in medium culture. Furthermore, it is demonstrated that existence of Cumulus Oocyte Complexes (COC) significantly showed better IVM, fertilization and evolution trend as compared to oocytes without cumulus cover or Denuded Oocytes (DO), especially in TCM199 plus BME and CYS. So that the change in GV stage oocytes to MII (maturation rate), fertilization rates or 2PN formation, and two cell embryos formation or blastocyst development rate in the treatment group with addition of BME & CYS and COC was statistically significant as compared to the DO group (p-value < 0.0001).

Conclusion: Both cellular and environmental factors could be important and involved in ART improvement.

Keywords: Cumulus oocyte complexes; Infertility; Oxidative stress; Reactive oxygen species.

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Figures

[Table/Fig-1]:
[Table/Fig-1]:
Mouse oocytes with ×300 magnification by light-microscope. a: Cumulus oocyte complexes (COC); b: Denuded oocytes (DO).
[Table/Fig-2]:
[Table/Fig-2]:
Maturation, fertilization, cleavage rates and blastocyst development percentage differences of mouse GV oocytes in presence or absence of BME and CYS. COC: Cumulus Oocyte Complex; DO: Denuded Oocytes, M II: Maturation Rate, GVBD: Germinal Vesicle Breakdown
[Table/Fig-5]:
[Table/Fig-5]:
Evolution trend of the Cumulus Oocyte Complex (COC) and Denuded Oocytes (DO) maturation inTCM199 medium. A: Metaphase II (MII); B: Second polar body (after fertilization); C: Fertilization (2PN); D: Two-cell embryos; E: Four-cell embryos; F: Eight-cell embryos; G: Blastocyst.
See this image and copyright information in PMC

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