Background: Typical difficulties encountered during in vitro fertilization (IVF) to produce embryos in pigs include poor pronucleus formation and poor-quality fertilized embryos because of high polysperm invasion. In this study, we evaluated the effects of supplementation with apple seed extract (ASE) and coculture systems on porcine in vitro-fertilized embryo culture. Methods: Slaughterhouse-derived ovaries were used to obtain cumulus-oocyte complexes (COCs). COCs were conventionally used to perform IVF. We examined the differences in apoptosis and metabolism during development following addition of ASE to normal culture and coculture systems. Matrix metalloproteinases (MMPs), cell development-related factors, and apoptotic proteins were compared in porcine embryos produced under different conditions. Results: The expression of genes related to insulin-like growth factor (IGF) signaling was increased in the coculture system. In the ASE group, early apoptosis and necrosis were reduced in fertilized embryos and the late survival rate increased. Supplementation of the coculture system with ASE led to increased expression of BCL-2 and decreased expression of Casp-3 in the cytoplasm, thereby lowering the apoptosis rate and inducing MMP expression. In addition, compared with the extract-supplemented group in normal culture, the activity of MMP-2 decreased in the coculture system supplemented with ASE, activity of MMP-9 increased, and the expression of dynactin p62 and BrdU in the cytoplasm was higher than that in the other groups. Conclusions: The coculture system increased the activity of the embryonic cytoplasm compared with the non-coculture system. Supplementation with ASE may induce cell activity and inhibit the expression of apoptotic factors.
Gold kiwifruit was fermented with Lactobacillus plantarum CK10 derived from kimchi and the fermented products were extracted with ethanol at various fermentation time-points The bacterial cellular density, total titratable acidity, total polyphenol content (TPC), and total flavonoid content (TFC) increased during fermentation, while pH values and total soluble solids decreased. Levels of TPC and TFC were highest after five days, at 1.21±0.13 mg GAE/g dry weight and 0.36±0.04 mg RE/g dry weight, respectively. The antioxidant activities of the fermented gold kiwifruit were analyzed using Fe2+ chelating activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH)-, and 2,2'-azino-bis(3- ethylbenzothiazoline-6-sulphonic acid) (ABTS)- radical scavenging activities, and superoxide dismutase (SOD)-like activity. The Fe2+ chelating activity of gold kiwifruit (125-500 μg/mL) peaked after five days of fermentation at 38.40-78.47%. The DPPH radical- scavenging activity and SOD-like activity were somewhat higher after seven days of fermentation (36.01-86.81% and 54.79-93.83% at 2.5-10.0 mg/mL concentration of samples, respectively). On the other hand, the ABTS radical- scavenging activity of fermented gold kiwifruit was similar to that of the non-fermented form. The polyphenol and flavonoid contents were significantly correlated with the antioxidant activity. In conclusion, our results suggest that TPC, TFC, and antioxidant activity were increased after five and seven days of fermentation, respectively. Therefore, fermented gold kiwifruit with its increased antioxidant activity could be useful in the development of functional foods.