Improved developmental competence in embryos treated with lycopene during in vitro culture system

Abstract

In vitro embryo development remains suboptimal compared to in vivo development due to the challenge from various stressors associated with in vitro culturing of oocytes. When 0.2 μM lycopene was added to oocyte in vitro maturation and embryo culture media, to assess its antioxidant effects on embryo development, we observed a significant (p < 0.05) increase in cleavage and blastocyst development rates compared to the corresponding controls (84.3 ± 0.6% vs. 73.1 ± 1.9% and 41.0 ± 1.4% vs. 33.4 ± 0.7%, respectively). Lycopene also significantly reduced (p < 0.05) intracellular reactive oxygen species concentrations in oocytes and blastocysts, whereas lipid peroxidation and mitochondrial activity increased compared to control conditions. The number of apoptotic nuclei was significantly reduced in the lycopene-treated compared to the control group (1.7 ± 0.1 vs. 4.7 ± 0.3), and the quantity of cells in the trophectoderm (207.1 ± 1.6 vs. 171.3 ± 1.0, respectively) and inner cell mass (41.9 ± 0.4 vs. 36.7 ± 0.4, respectively) was higher following treatment-although the inner cell mass-to-trophectoderm ratio was unchanged (1:3.3 vs. 1:3.4 for lycopene vs. control, respectively). Lycopene supplementation also significantly (p < 0.05) attenuated expression of IKBKB (Inhibitor of nuclear factor kappa B kinase, subunit beta) and reduced Caspase 9 and Caspase 3 protein abundance, while up-regulating GDF9 (Growth and differentiation factor 9), BMP15 (Bone morphogenetic protein 15), SOD2 (Superoxide dismutase 2), NDUFA2 (NADH dehydrogenase), ACADL (Acyl-CoA dehydrogenase, long chain), and ACSL3 (Acyl-CoA synthetase 3, long-chain membrane 3) transcription compared to control. Therefore, co-culturing with lycopene during oocyte maturation improved bovine embryo developmental potential during in vitro culture by improving embryonic resilience to stress.

The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos

“…#11360-070). The oocytes selected were allowed to undergo in vitro maturation at 38.5 °C in a humidified atmosphere of 5% CO 2 in air for 22–24 h [55].…”

“…The IVM matured oocytes were fertilized in vitro with frozen–thawed bovine sperm, as described previously [55]. In brief, the sperm were thawed at 38.0 °C for 1 min and diluted in D-PBS.…”

PTPN11 (SHP2) Is Indispensable for Growth Factors and Cytokine Signal Transduction During Bovine Oocyte Maturation and Blastocyst Development

“…IVM COCs were fertilized with frozen–thawed bovine sperm, as previously described [38]. In brief, semen was thawed at 39.0 °C for 1 min and the sperm were washed in D-PBS, followed by centrifugation at 750 × g for 5 min at room temperature.…”

“…Immunofluorescence staining was performed as previously discussed [38,39]. Briefly, oocytes or blastocysts were fixed in 4% ( v / v ) paraformaldehyde prepared in 1 M phosphate-buffered saline (PBS) and preserved at 4 °C.…”

“…ROS was measured through 2,7,dichlorodihydrofluorescein diacetate (H 2 DCFDA) as described previously [38]. In brief, the live mature (MII) oocytes (30 to 35 used per each group/experiment) or day-8 blastocysts (10 to 15 used per each group/experiment) were incubated in 1 × PBS containing 10 μM of H 2 DCFDA for 30 min in a humidified atmosphere of 5% ( v / v ) CO 2 in air at 38.5 °C.…”