Synergistic Effects of Glutathione and beta-Mercaptoethanol Treatment During In Vitro Maturation of Porcine Oocytes on Early Embryonic Development in a Culture System Supplemented with L-cysteine

Abstract

Various methods have been used to remove reactive oxygen species (ROS) generated from in vitro culture (IVC) conditions that can cause cell injury or death, including the application of low oxygen (O-2) tension and the addition of antioxidants The beneficial effects of antioxidants and O-2 tension on IVC of porcine embryos, however, are controversial among researchers In this study, we sought to determine the effects and optimal concentrations of antioxidants for the development of porcine embryos in an IVC system Specifically, we examined the synergistic effects of antioxidants on development to the blastocyst stage in a culture system supplemented with L-cysteine during IVM Of the antioxidants tested (melatonin, glutathione (GSH), beta-mercaptoethanol (beta-ME), N-acetylcysteine (NAC) and dithiothreitol (DTT)), addition of GSH (1 mM) or beta-ME (25 mu M) significantly increased development to the blastocyst stage compared with the controls without antioxidant treatment (22 2 +/- 4 2% for 1 mM GSH, 25 9 +/- 2 2% for 25 mu M beta-ME and 12-13% for the control, P<0 05) In addition, the mean cell number per blastocyst was increased by approximately 1 7 fold in the presence of GSH or beta-ME These GSH- and beta-ME-induced increases m development to the blastocyst stage and total cell number, however, were not mimicked by melatonin, NAC or DTT, all of which are ROS scavengers The combination of GSH or beta-ME with L-cysteine significantly reduced high O-2 tension-induced ROS production (P<0 05) These results suggest that a combination of 1 mM GSH or 25 mu M beta-ME with 1 mM L-cysteine could be used for production of high quality porcine blastocysts in IVC systems