Cysteamine enhances in vitro development of porcine oocytes matured and fertilized in vitro.

Abstract

Most porcine oocytes matured and fertilized in vitro fail to develop normally due to abnormal fertilization. The aim of this study was to determine the effect of cysteamine on pronuclear formation and developmental competence in pig embryos produced in vitro. Follicular oocytes were matured in vitro with or without cysteamine in Medium 199 supplemented with sodium pyruvate, FSH, LH, 17 beta-estradiol, antibiotics, and 25% porcine follicular fluid. Matured oocytes were then inseminated for 10 h. In the first experiment, pronuclear formation was assessed immediately after the insemination period. The addition of 50 microM or 500 microM cysteamine to the maturation medium increased male pronuclear development, resulting in a higher proportion of oocytes with synchronously formed male and female pronuclei. The frequency of synchronous pronuclear formation was increased in monospermic oocytes from 10% in the control group to 43% (p < 0.001) and 45% (p < 0.001), respectively, and in polyspermic oocytes from 43% in the control group to 68% (p < 0.05) and 75% (p < 0.001), respectively. In the second experiment, development of the in-vitro produced embryos was assessed after 7 days of culture in vitro. The addition 500 microM cysteamine to the maturation medium increased the percentage of cleaving embryos developing to the 8-cell (37% vs. 16%; p < 0.001), morula (19% vs. 6%; p < 0.001), and blastocyst (12% vs. 1%; p < 0.001) stages. These results demonstrate that the addition of cysteamine to maturation medium increases synchronous pronuclear formation and normal embryonic development in porcine oocytes matured and fertilized in vitro.