The influence of ascorbic acid on in vitro maturation of canine oocytes

Abstract

It is known that L-ascorbic acid (vitamin C) can modulate many biochemical processes intracellularly or extracellularly as antioxidant. The aim of the present study was to examine the effects of media supplementation with ascorbic acid on canine oocyte meiotic maturation, viability and the cumulus cell expansion. Various concentrations of ascorbic acid supplemented in in vitro maturation (IVM) media were tested. Canine oocyte was exposed to different levels of ascorbic acid (0, 50, 150, 250, 500, 750µM). Cumulus expansion, meiotic maturation and degeneration of oocytes were assessed 72 h after in vitro culture. As results, on the group treated with 250µM ascorbic acid was a significant difference compared to the control group on nuclear maturation in stages metaphase I (MI) and metaphase II (MII) (26.98% vs. 6.00%). The groups treated with 50, 150, 250, 500µM had an increase in stage (GVBD), and a significant decrease of degenerate-undefined oocytes compared with the control (23.31%, 18.85%, 13.41% vs 40.80). Concentration 750µM had similar effect to that in the control group. The groups treated with 50, 150, 250, 500µM had an increase in meiosis resumption (GVBD), metaphase I (MI) and metaphase II (MII) with the best result in the group treated with 250 µM ascorbic acid.

Effect of antioxidant ascorbic acid on in vitro maturation of Caprine oocytes under normal and elevated temperatures

“…Nuclear maturation status achieved with addition of ascorbic acid is comparable to findings of Pernes et al, (2016) who found that ascorbic acid increased the meiotic resumption of canine oocytes and higher proportions of oocytes reached MI and MII stage. Significant improvement in maturation rate of MII oocytes with ascorbic acid supplemented media were reported by Dorra et al, (2012) and Nadri et al, (2009) in rabbit and mice oocytes, respectively.…”

“…It inhibited the apoptosis in bovine granulosa cells and murine cumulus oocyte complexes (Tilly and Tilly, 1995;Eppig et al, 2000;Murray et al, 2001), reduced intracellular levels of reactive oxygen species (Castillo-Martín et al, 2014b) and heat shock protein 70 (HSPA1A) (Castillo-Martín et al, 2014a). As antioxidant supplement, ascorbic acid has many beneficial effects including improvement of nuclear maturation and developmental competence of oocytes matured in vitro (Pernes et al, 2016). The role of ascorbic acid as antioxidant on in vitro maturation of oocytes is documented by various authors in literature; however the studies on the effect of ascorbic acid on in vitro maturation of oocytes under heat stress are very less.…”

Utilization of Rosmarinic and Ascorbic Acids for Maturation Culture Media in Order to Increase Sow Oocyte Quality Prior to IVF

“…In vitro embryo production is necessary in order to obtain high quality embryos that are used in reproductive biotechnology, as well as in research [ 4 , 10 , 12 ].…”

“…In the practice of IVF ascorbic acid has often been used in different concentrations as a supplement in in vitro culture systems for embryos but also for the maturation of domestic cat oocytes [ 9 ], buffalo and bovine oocytes [ 10 , 11 ], canine oocytes [ 12 ] or pig oocytes [ 8 ]. The better results in their studies were with 50 µg/mL ascorbic acid in bovine oocytes [ 11 ], 50 µM for buffalo oocytes [ 10 ], 250 µM for canine oocytes [ 12 ], or 250 µM for pig oocytes [ 8 ].…”

“…In the practice of IVF ascorbic acid has often been used in different concentrations as a supplement in in vitro culture systems for embryos but also for the maturation of domestic cat oocytes [ 9 ], buffalo and bovine oocytes [ 10 , 11 ], canine oocytes [ 12 ] or pig oocytes [ 8 ]. The better results in their studies were with 50 µg/mL ascorbic acid in bovine oocytes [ 11 ], 50 µM for buffalo oocytes [ 10 ], 250 µM for canine oocytes [ 12 ], or 250 µM for pig oocytes [ 8 ]. In sperm cryopreservation protocols, ascorbic acid has given really good results due to its antioxidant capacity, especially with a concentration of 3 mg/mL, or a concentration of 5 mM [ 13 ] for bull semen or a concentration of 600 µM in human semen during vitrification [ 14 ].…”