Ascorbic Acid Improves the Developmental Competence of Porcine Oocytes After Parthenogenetic Activation and Somatic Cell Nuclear Transplantation

Abstract

In this study, a dose-response assessment was performed to understand the relation between supplementation of media with L-ascorbic acid or vitamin C and porcine oocyte maturation and the in vitro development of parthenotes (PA) and handmade cloned (HMC) embryos. Various concentrations (0, 25, 50 and 100 µg/ml) of vitamin C supplemented in in vitro maturation (IVM) and culture (IVC) media were tested. None of these vitamin C additions affected nuclear maturation of oocytes, yet supplementation at 50 µg/ml led to significantly increased intracellular glutathione (GSH) levels and reduced reactive oxygen species (ROS). When cultured in IVM- and/or IVC-supplemented media, the group supplemented with 50 µg/ml of vitamin C showed improved cleavage rates, blastocyst rates and total cell numbers per blastocyst (P<0.05) compared with other groups (control, 25 µg/ml and 100 µg/ml). In contrast, supplementation with 50 µg/ml vitamin C decreased (P<0.05) the apoptosis index as compared with the groups supplemented with 100 µg/ml. In addition, even with a lower blastocyst rate to start with (37.6 vs. 50.3%, P<0.05), supplementation of HMC embryos with vitamin C ameliorated their blastocyst quality to the extent of PA embryos as indicated by their total cell numbers (61.2 vs. 59.1). Taken together, an optimized concentration of vitamin C supplementation in the medium not only improves blastocyst rates and total cell numbers but also reduces apoptotic indices, whereas overdosages compromise various aspects of the development of parthenotes and cloned porcine embryos.

Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice

“…Intracellular GSH levels were determined by previously reported methods [14,32]. Briefly, cell tracker blue CMF2HC (4-chloromethyl-6, 8-difluoro-7-hydroxycoumarin; Molecular probe, Life technology) was used to detect intracellular GSH level by blue fluorescence.…”

Effect of using ascorbic acid and cysteamine supplementation on in-vitro development of buffalo embryos

“…It was shown that the optimum concentration of vitamin C was 50 µg/mL during IVM of porcine oocytes [20], this concentration not only improved blastocyst rates, total cell numbers but also, reduced apoptotic indices and the high concentration (100 µg/mL) of vitamin C induced high apoptotic index. Miclea [21] and Huang [22] mentioned that the treatment with 50 µg/mL vitamin C resulted in increasing pregnancy rate and enhancing the development somatic cell nuclear transfer embryos in pigs.…”

“…The beneficial effect of vitamin C addition might be a scribed to an improved the culture condition and oocytes cytoplasmic maturation by reduction the intra cellular oxidative status for embryo development [7,20]. Moreover, Eppig [24] and Wang [6] mentioned that the addition of ascorbic acid into the culture media prevented follicular apoptosis in rat and mouse follicles, and also improved the blastocyst production.…”

Relationship Between Follicular Fluid and Serum Levels of Vitamin C and Oocyte Morphology and Embryo Quality in Patients Undergoing In Vitro Fertilization

“…But supplementation of 50 μg/mL led to significantly increased intracellular glutathione levels and reduced ROS and improved cleavage rates, blastocyst rates and total cell numbers per blastocyst. Also supplementation with 50 μg/mL vitamin C decreased the apoptosis index as compared with the groups supplemented with 100 μg/mL (19). Barzegari Firouzabadi's study which investigated the effects of ascorbic acid on the IVM of mouse's follicles and enclosed oocytes mentioned that ascorbic acid could increase survival rate but did not affect diameter, GV breakdown, and oocyte maturation rates (22).…”