Influence of commercially available follicle stimulating hormone on the in vitro maturation of bovine oocytes

Abstract

The work aimed (Experiment I) to compare commercial representations of porcine follicle stimulating hormone (FSH, Pluset® vs. Folltropin®) in concentration (10 μg/mL) and time (24 h) standard (more used in protocols of in vitro maturation, IVM); (Experiment II) to evaluate the best incubation time (6 h vs. 16 h vs. 24 h) and, (Experiment III) to analyze varying concentrations (1.0 μg/mL vs. 2.5 μg/mL vs. 10.0 μg/mL) of representations of FSH on the IVM of bovine oocytes. Thus, oocytes were recovered and submitted to IVM under appropriate conditions. After the IVM, oocytes were evaluated for expansion of cumulus cells (CCs), presence of the first polar body (1PB) and metaphase plate (MII). All the data were analyzed by the Fisher exact test (P < 0.05). Initially, in Experiment I, no difference (P > 0.05) was observed in maturation rates of the oocytes incubated with FSH Pluset® or Folltropin®, assessed by expansion of CCs (97.6% vs. 94.3%), presence of 1PB (76.6% vs. 69.4%) and MII (70.0% vs. 68.6%). In Experiment II, when the incubation time with FSH was evaluated, both Pluset® as Folltropin® showed lower rate of expansion of CCs when they were present only in the first 6 h of IVM. As for presence of 1PB, differences were observed in relation to Pluset® while Folltropin® showed similar results in all incubation times. Regarding the MII, no difference was observed between the incubation times with FSH Pluset® and Folltropin®. In Experiment III, no difference was observed in the expansion of CCs, presence of 1PB and MII for concentrations evaluated FSH Pluset® and Folltropin®. Therefore, the FSH Pluset® and Folltropin® have the same efficiency in IVM of bovine oocytes. Regarding the incubation time, it is recommended to maintain FSH (Pluset® or Folltropin®) throughout the period of IVM, since there was no difference in the results of presence of MII. Furthermore, the concentration of 1.0 μg/mL of FSH Pluset® and Folltropin® is as effective as 10 μg/mL and can therefore be used for IVM of oocytes. Key words: Cattle. Gonadotropin. In vitro production of embryos. Oocytes. ResumoO trabalho objetivou (Experimento I) comparar representações comerciais do hormônio folículo estimulante suíno (FSH, Pluset® vs. Folltropin®) na concentração (10 μg/mL) e tempo (24 h) padrão (mais utilizados nos protocolos de maturação in vitro, MIV); (Experimento II) avaliar o melhor tempo de incubação (6 h vs. 16 h vs. 24 h) e, (Experimento III) analisar concentrações variáveis (1,0 μg/mL vs. 2,5 μg/mL vs. 10,0 μg/mL) das representações de FSH sobre a MIV de oócitos bovinos. Para tanto, oócitos foram recuperados e submetidos a MIV em condições adequadas. Após a MIV, oócitos foram avaliados quanto à expansão das células do cumulus (CCs), presença do primeiro corpúsculo polar (1CP) e placa metafásica (MII). Todos os dados foram analisados pelo teste exato de Fisher (P < 0,05).

Use of natural antioxidants in in vitro mammalian embryo production

“…Moretti et al (2012) , and increased GSH concentration when they used 10.0 µg/mL of this antioxidant. Moreover, when crocin was added to the medium of bovine sperm capacitation, satisfactory results obtained both in sperm quality parameters (motility, intact acrosome, acrosome reaction, and DNA fragmentation) as well as in IVF and IVD using these gametes (SAPANIDOU et al, 2015). In this study, the blastocyst rate (54.2%) was significantly higher with the addition of 1.0 mM of crocin in sperm capacitation compared to the control without antioxidant (37.5%) (SAPANIDOU et al, 2015).…”

“…Moreover, when crocin was added to the medium of bovine sperm capacitation, satisfactory results obtained both in sperm quality parameters (motility, intact acrosome, acrosome reaction, and DNA fragmentation) as well as in IVF and IVD using these gametes (SAPANIDOU et al, 2015). In this study, the blastocyst rate (54.2%) was significantly higher with the addition of 1.0 mM of crocin in sperm capacitation compared to the control without antioxidant (37.5%) (SAPANIDOU et al, 2015). Crocetin was used in the IVD medium, where a concentration of 1.0 μM could significantly improve the blastocyst rate (46.1%) and embryo quality by decreasing the percentage of apoptotic cells and increasing cryotolerance (ZULLO et al, 2016b).…”

“…In contrast, isolated substances allow greater control over the bioactive that will be tested. For this, chromatographic techniques, especially high performance liquid chromatography, and immunoassays have been used (SASIDHARAN et al, 2011).…”

“…These media must provide all the nutrients and factors necessary for the development of gametes and embryos, as well as maintain the balance of molecules that can harm the system (CROCOMO et al, 2012;SANTOS et al, 2017). In this regard, one of the main factors that can influence the success of IVEP is oxidative stress caused by the accumulation of reactive oxygen species (ROS) that impair the integrity of cells and their macromolecules thus interfering with in vitro development (ROCHA-FRIGONI et al, 2015).…”

Influence of different protein supplements on the recovery and in vitro maturation of bovine oocytes

“…These initial stages of oocyte recovery and maturation are of great importance for better results, as the quality of cumulus-oocyte complexes (COCs) obtained determines the later stages of the technique (Santos et al, 2016;Santos et al, 2017). Therefore, to ensure nutrition and energy support during in vitro oocyte development, the media requires supplementation with external protein sources.…”