l-carnitine supplementation during in vitro culture regulates oxidative stress in embryos from bovine aged oocytes

2020 ◽  
Vol 143 ◽  
pp. 64-73 ◽  
Author(s):  
Wenjie Jiang ◽  
Yinghua Li ◽  
Yuhan Zhao ◽  
Qingshan Gao ◽  
Qingguo Jin ◽  
...  
2008 ◽  
Vol 28 (1) ◽  
pp. 36-45 ◽  
Author(s):  
Biljana Balen ◽  
Mirta Tkalec ◽  
Dubravko Pavoković ◽  
Branka Pevalek-Kozlina ◽  
Marijana Krsnik-Rasol

2008 ◽  
Vol 104 (2-4) ◽  
pp. 132-142 ◽  
Author(s):  
Geórgia Assis Corrêa ◽  
Rodolfo Rumpf ◽  
Tatiane Carmo Duarte Mundim ◽  
Mauricio Machaim Franco ◽  
Margot Alves Nunes Dode

2019 ◽  
Vol 7 (2) ◽  
pp. 66-76
Author(s):  
Katarzyna Stefańska ◽  
Sandra Knap ◽  
Magdalena Kulus ◽  
Ievgenia Kocherova ◽  
Piotr Celichowski ◽  
...  

AbstractOxygen metabolism is crucial in establishing successful pregnancy, since excessive amount of reactive oxygen species (ROS) may exert deleterious effects on the developing embryo. There are several defense mechanisms against oxidative stress in the female reproductive tract, including production of antioxidant enzymes by oviductal epithelial cells (OECs). Undoubtedly, OECs play major part in female fertility and may also serve as an in vitro model of the oviduct. Therefore, the aim of this study was to investigate the expression of genes involved in oxygen metabolism. We have isolated OECs from oviducts of crossbred gilts (n=45) and maintained their in vitro culture for 30 days, collecting their RNA at days 1, 7, 15 and 30. The gene expression was determined with the use of Affymetrix® Porcine Gene 1.1 ST Array Strip. Our results revealed 166 differentially expressed genes belonging to four ontology groups: „cellular response to oxidative stress”, “cellular response to oxygen-containing compound”, “cellular response to oxygen levels” and “cellular response to reactive oxygen species”, most of which are also involved in other major processes in the organism. However, our findings provide a valuable insight into porcine reproductive biology and may be utilized in optimization of assisted reproduction techniques.Running title: Genes involved in oxygen metabolism in oviductal epithelial cells


2020 ◽  
Vol 21 (16) ◽  
pp. 5601
Author(s):  
Diego F. Carrillo-González ◽  
Nélida Rodríguez-Osorio ◽  
Charles R. Long ◽  
Neil A. Vásquez-Araque ◽  
Juan G. Maldonado-Estrada

l-carnitine is a potent antioxidant used for in vitro culture systems. Controversial results have been reported using l-carnitine in culture medium at different stages of in vitro bovine embryo production. Cumulus-oocyte complexes (n = 843) were in vitro-fertilized and cultured and added (treatment group) or not added (control group) with l-carnitine. At day three of culture, each group was subdivided into two subgroups receiving no l-carnitine (group 1), 3.8 mM l-carnitine added during in vitro maturation (group 2), 1.5 mM added during the in vitro culture (group 3), and 3.8 mM and 1.5 mM added during the maturation and culture, respectively (group 4). At day 8, blastocyst embryos were examined for mitochondrial activity, the presence of lipid droplets, total cell number, gene expression, and cryotolerance by vitrification. The data were analyzed with a one-way analysis of variance. l-carnitine added in the late in vitro culture significantly reduced mitochondrial activity and lipid content, and upregulated ifn-τ and ptgs2 gene expression compared to controls (p < 0.05). l-carnitine supplementation did not significantly affect the embryo rate production or survival rate after vitrification and warming (p > 0.05). l-carnitine supplementation significantly improved embryo potential to develop viable pregnancies in agreement with a study reporting improved pregnancy rates.


2013 ◽  
Vol 25 (1) ◽  
pp. 214 ◽  
Author(s):  
L. Boccia ◽  
M. De Blasi ◽  
G. Zullo ◽  
V. Longobardi ◽  
D. Vecchio ◽  
...  

In buffalo, in vitro embryo production (IVEP) technology is the best tool to improve the genetic merit through the maternal lineage. A major limitation of IVEP technology in buffalo species is the poor cryotolerance of the embryos, likely due to their high lipid content (Gasparrini 2002 Theriogenology 57, 237–256). It was previously demonstrated that supplementing bovine culture media with L-carnitine, a cofactor of β-oxidation, improves in vitro embryo development (Sutton-McDowall et al. 2012 Theriogenology 77, 1632–1641). The aim of this work was to evaluate whether L-carnitine supplementation during in vitro culture (IVC) improves blastocyst development and cryotolerance of in vitro produced buffalo embryos. After a preliminary dose response trial, we selected the concentration of 0.25 mM for the experiment. Cumulus–oocytes complexes (n = 288, over 4 replicates), recovered from slaughtered animals, were matured and fertilized in vitro according to our standard procedures (Gasparrini et al. 2006 Theriogenology 65, 275–287). On Day 1 (Day 0 = IVF), zygotes were cultured in SOF supplemented with 8 mg mL–1 BSA, in the absence (control, n = 143) or presence of 0.25 mM L-carnitine (n = 145). In vitro culture was carried out at 38.5°C under 5% CO2, 7% O2, and 88% N2. Cleavage rate was evaluated on Day 5, when the cleaved embryos were transferred into fresh medium for further 2 days. On Day 7 after IVF, embryo outcome was assessed and all the embryos were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide (DMSO), and 0.5 M sucrose (De Rosa et al. 2007 Ital. J. Anim. Sci. 6(Suppl 2), 747–750). The resistance to cryopreservation was evaluated by assessing the survival rate, on the basis of morphological criteria, after 24 h culture. Data were analyzed by chi-square test. No differences were found in cleavage rates between the control (81.5%) and the L-carnitine group (78.8%). The blastocyst yields (calculated in relation to the cleaved embryos) were not significantly influenced by the L-carnitine treatment (40.2 and 52.9%, in the control and the L-carnitine groups, respectively). However, buffalo embryos cultured in the presence of L-carnitine showed an increased resistance to cryopreservation, as indicated by the higher survival rates recorded after 24 h culture (78.7 and 96.4%, in the control and the L-carnitine groups, respectively; P < 0.01). In conclusion, these results demonstrated that L-carnitine supplementation of culture medium improves the resistance to cryopreservation of in vitro produced buffalo embryos. We speculate that the increased cryotolerance observed in the presence of L-carnitine may be due to a better utilization of the endogenous lipid stores, resulting in improved embryo quality.


2015 ◽  
Vol 27 (1) ◽  
pp. 160
Author(s):  
G. Zullo ◽  
A. Salzano ◽  
G. Bifulco ◽  
V. Longobardi ◽  
G. Albero ◽  
...  

It is known that in vitro mammalian embryo development is negatively affected by the increased oxidative stress occurring under culture conditions. The oxidative damage of cell components via reactive oxygen species interferes with proper cell function. Buffalo embryos are particularly sensitive to oxidative stress because of their high lipid content (Boni et al. 1992 Acta Med. Vet. 38, 153–161). l-Ergothioneine (LE) is a powerful scavenger of hydroxyl radicals (OH) and an inhibitor of iron or copper ion-dependent generation of OH from hydrogen peroxide (H2O2). The aim of this study was to evaluate whether enriching the in vitro-culture medium with LE improves in vitro embryo production efficiency in buffalo. Abattoir-derived buffalo oocytes (n = 854, over 6 replicates) were in vitro matured and fertilized according to standard procedures (Gasparrini et al. 2006 Theriogenology 65, 275–287). Twenty hours after IVF presumptive zygotes were cultured in SOFaa supplemented by 8 mg mL–1 BSA in a controlled gas atmosphere consisting of 5% CO2, 7% O2, 88% N2, in humidified air, at 38.5°C with 0 (control; n = 214), 0.05 mM LE (n = 217), 0.1 mM LE (n = 204), and 1 mM LE (n = 219). Cleavage rate was assessed at the time of change of culture (Day 5) and the cleaved elements were cultured for a further 2 days. The embryos obtained by the end of culture, i.e. on Day 7 post-IVF, were scored for quality, based on morphological criteria, and for developmental stage, as previously described (Robertson, Nelson 2010 Manual of the International Embryo Transfer Society 86–105). The percentages of total transferable embryos and Grade 1 and 2 blastocysts in relation to cleaved oocytes were recorded. Because the chronology of development is known to be one of the most reliable parameters for assessing quality, the percentage of fast-developing embryos, i.e. hatched and expanded blastocysts, was also recorded. Data were analysed by Chi-squared test. Cleavage rate was not affected by the treatment (71.4, 66.8, 68.7, and 63.0%, respectively, with 0, 0.05, 0.1, and 1 mM LE). The total embryo output increased in groups supplemented with 0.05 and 0.1 mM LE (31.3, 42.2, 43.8, and 21.7%, respectively, with 0, 0.05, 0.1, and 1 mM LE; P < 0.05). However, the enrichment of in vitro culture with 0.1 mM LE also increased the percentage of Grade 1 and 2 blastocysts compared with the control and to 1 mM LE (21.6, 30.9, 33.9, and 21.7%, respectively, with 0, 0.05, 0.1, and 1 mM LE; P < 0.05). Likewise, 0.1 mM LE gave higher percentages of fast developing embryos than the control and 1 mM LE groups. In conclusion, these results demonstrated a beneficial effect of LE during culture on buffalo in vitro embryo development. The dose response trial indicated that the optimal concentration is 0.1 mM that also influenced the chronology of development and hence embryo viability.


2021 ◽  
Author(s):  
Jing Chen ◽  
Evgenia Isachenko ◽  
Wanxue Wang ◽  
Xinxin Du ◽  
Mengying Wang ◽  
...  

Abstract Background: This study aims to evaluate the effectiveness of Tumor Dissociation Enzyme (TDE) for the isolation of cryopreserved human ovarian cortex tissues, proposing an optimized applicable for artificial ovary.Methods: This is a prospective experimental study. We present a comparative analysis among the outputs of follicles isolated from cryopreserved ovarian biopsies of ten young women undergoing laparoscopy. Follicles were isolated by Tumor Dissociation Enzyme (TDE) or Liberase Dispase High (DH) enzymatic digestion. Follicles were assessed by evaluating the number, viability, morphology, oxidative stress. Moreover, the follicle growth and viability were analyzed after eight days of in vitro culture (IVC) between each protocol.Results: The recovery rate of follicles in Group 1 was significantly higher by 78 follicles compare to Group 2 (p < 0.05), while the difference in weight and volume of biopsies in both groups was no significant (p > 0.05). Group 1 allowed to isolate more primordial follicles (p < 0.05) and smaller diameter of follicles (p < 0.01) than Group 2. Group 1 allowed to isolate 7% more of bright red follicles than Group 2 (p < 0.01). We also found that Group 1 had isolated a significantly higher percent of viable follicles (p < 0.05), more morphologically normal follicles (p < 0.05), and lower oxidative stress levels compared with Group 2 (p < 0.01). The velocity of follicles growth from Day 0 to Day 8 in Group 1 was significantly higher (p < 0.05) than in Group 2. The viability of follicles on Day 8 of in vitro culture in Group 1 was significantly higher than Group 2 (p < 0.05).Conclusions: TDE treatment can be an alternative for Liberase DH, allows the isolation of highly viable follicles from the cryopreserved human ovarian cortex, with an intact morphology and low oxidative stress, and with high proliferation potential after culture in vitro.


2019 ◽  
Vol 8 (6) ◽  
pp. 289 ◽  
Author(s):  
DiegoF Carrillo-Gonzalez ◽  
Nélida Rodríguez-Osorio ◽  
CharlesR Long ◽  
NeilA Vásquez-Araque ◽  
JuanG Maldonado-Estrada

Planta Medica ◽  
2010 ◽  
Vol 76 (12) ◽  
Author(s):  
K Graikou ◽  
H Damianakos ◽  
K Syklowska-Baranek ◽  
A Pietrosiuk ◽  
M Jeziorek ◽  
...  

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