Effects of l‐Ascorbic Acid, α‐Tocopherol and Co‐culture on In Vitro Developmental Potential of Porcine Cumulus Cells Free Oocytes

Abstract

The maturation and developmental potential on cumulus-cell-free oocytes is of great importance theoretically and practically. The present study was to investigate the effects of l-ascorbic acid, alpha-tocopherol and co-culture on in vitro developmental potential of porcine denuded oocytes (DOs). Porcine DOs were cultured in maturation medium supplemented with vitamin C (0, 50, 100, 250, 500, 750 microM) and vitamin E (0, 10, 20, 50, 100, 250 microm), respectively. And they were also co-cultured with dispersed cumulus cells (group CCscoculture), intact cumulus cells oocyte complexes (COCs) (group COCscoculture), and COCs whose oocytes were removed (group OOXcoculture), respectively. After 44 h incubation, the maturation rates, cleavage rates and blastocyst rates after parthenogenetic activation in three experiments mentioned above were collected and analysed, respectively. L-Ascorbic acid promoted porcine DOs in vitro maturation and blastocyt development after parthenogenetic activation while alpha-tocopherol did not increase the in vitro maturation rates, but improved the blastocyst rate. None of the three co-culture manner promoted the in vitro maturation and the cleavage of porcine DOs after parthenogenetic activation, but all the co-culture manners improved the blastocyst rates. Both Vitamin C and E enhance the in vitro developmental potential of porcine DOs. Co-culture increases the developmental potential of porcine DOs.

Effects of oxygen concentration on in vitro maturation of canine oocytes in a chemically defined serum-free medium

“…Vitamin E in the presence of high oxygen was able to decrease degeneration (PZ0.008) but did not improve the percentage of oocytes in MII. The beneficial effect of vitamin E on oocyte maturation and embryo development was previously reported in animal species regardless of lipid content, including porcine cells (Tao et al 2010), which contain high lipid, or in ovine oocytes (Natarajan et al 2010), which contain relatively low lipid contents.…”

“…This may reflect the importance of energy supply during the prolonged period of oviductal travel and the maternal zygotic transition period (Guraya 1965, Luvoni et al 2005, Lopes et al 2010. However, presence of these abundant lipid droplets in ooplasm has an impeccable influence on predisposition of cumulus oocyte complexes (COCs) to oxidative stress by reactive oxygen species (ROS; Wakefield et al 2008, Whitaker & Knight 2008, Tao et al 2010. Oxygen concentration during the IVM culture period can contribute to the extent and velocity of this oxidative stress of which oocyte nuclear and cytoplasmic maturation and development pattern may perturb (Kim et al 2007, Whitaker & Knight 2008.…”

“…Vitamin E (a-tocopherol) as a lipid-soluble antioxidant has been proven to have beneficial effects on oocyte maturation and embryo development in pigs (Tao et al 2010) and alleviates the degeneration rates of bovine and ovine oocytes (Dalvit et al 2005, Natarajan et al 2010. This study investigated the impact of oxidative stress on oocyte nuclear maturation and degeneration.…”

Antioxidants rescue stressed embryos at a rate comparable with co-culturing of embryos with human umbilical cord mesenchymal cells

“…One mM β-ME and 50 μg/ml vitamin C were added to complete medium to prepare antioxidant-plus medium (Aopm) [13,16]. Fifty μl drops were loaded onto 60×15 mm culture dishes, overlaid by seven ml mineral oil, and incubated at 37°C in a humid atmosphere with 5% CO 2 for 24 h.…”

“…Vitamin C is a water-soluble vitamin and is the major antioxidant component in extracellular fluids [11,12]. Supplementation of bovine culture medium with vitamin C improved the porcine denuded oocyte maturation and blastocyst rate after parthenogenetic activation [13]. β-mercaptoethanol (β-ME) is also known as a low molecular weight antioxidant that is usually used in culture media, especially embryonic stem cell culture media [14,15].…”