Beneficial Effects of L-Carnitine Supplementation during IVM of Canine Oocytes on Their Nuclear Maturation and Development In Vitro

SIMPLE SUMMARY: In vitro production of canine embryos is a technique that can be used as a model to conserve endangered species and to establish efficient breeding systems for domestic dogs. However, compared with other species, the success rates of in vitro embryo production (IVEP) in dogs are low. L-Carnitine (LC) is a small water-soluble molecule; it plays an essential role in fatty acid metabolism and acts as a potent antioxidant. Various studies have reported the beneficial impacts of LC on IVEP in many mammalian species other than dogs. Therefore, these experiments investigated the effects of LC supplementation during in vitro maturation (IVM) on canine oocytes maturation, fertilization, and development in vitro. We show that the supplementation of IVM media with LC has positive impacts on oocyte maturation, fertilization, and preimplantation embryo development rates. We also demonstrate that 0.6 mg/mL LC is the most beneficial concentration to be used. It resulted in significantly higher maturation, fertilization, and embryo developmental rates than the control and other LC concentrations. These outcomes are essential for refining the IVM conditions that can advance the efficiency of assisted reproductive technologies (ARTs) in dogs. ABSTRACT: This study aimed to investigate the effect of L-Carnitine (LC) supplementation during in vitro maturation (IVM) of canine oocytes on nuclear maturation, fertilization status, and preimplantation development. Cumulus–oocyte complexes (COCs) collected from the ovaries of ovariohysterectomized female dogs were matured in vitro for 72 h in a TCM-199 medium supplemented with (0.1, 0.3, 0.6, 1.0, or 2.0 mg/mL) or without (0.0 mg/mL) LC. Matured oocytes were fertilized in vitro with frozen–thawed spermatozoa, and zygotes were cultured in a SOF medium for 7 days. IVM rates were higher (p ≤ 0.05) in 0.3 and 0.6 mg/mL LC supplemented groups than in the control (0.0 mg/mL LC) and other LC groups. Fertilization (18 h postinsemination (pi)) and cleavage (2–16-cell stage at day 3 pi) rates were higher (p ≤ 0.05) in the 0.6 mg/mL LC group than in the control and 0.1, 1.0, and 2 mg/mL LC supplemented groups. Interestingly, 4.5% of fertilized oocytes developed to morula (day 5 pi) in the 0.6 mg/mL LC group, which was higher (p ≤ 0.05) than those developed in the 0.3 mg/mL group (1.0%). No cleaved embryos developed to morula in other groups. In conclusion, LC supplementation at 0.6 mg/mL during IVM of canine oocytes improved their maturation, fertilization, and preimplantation embryo development rates following IVF and in vitro culture (IVC).