Effect of red propolis extract isolated or encapsulated in nanoparticles on the in vitro culture of sheep preantral follicle: Impacts on antrum formation, mitochondrial activity and glutathione levels

2018 ◽  
Vol 54 (1) ◽  
pp. 31-38 ◽  
Author(s):  
Tiago S. Nascimento ◽  
Isabelle S. M. Silva ◽  
Maria Cecília M. A. Alves ◽  
Bruna B. Gouveia ◽  
Lara Mariane R. Barbosa ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Mitochondrial Activity ◽  
Preantral Follicle ◽  
Propolis Extract ◽  
Red Propolis Extract

83 VITAMIN K2 SUPPLEMENTATION IMPROVES BLASTOCYST RATE BY RECOVERY OF MITOCHONDRIA IN IN VITRO-CULTURED BOVINE EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 155
Author(s):  
L. Baldoceda ◽  
C. Vigneault ◽  
P. Blondin ◽  
C. Robert
Keyword(s):  
In Vitro Culture ◽  
Fluorescent Microscopy ◽  
Vitamin K2 ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Rate ◽  
Mitotracker Red

Mitochondria play an important role during early mammalian embryo development through their diverse cellular functions, in particular creating balance between production of ATP by electron transport chain and oxidative stress. Embryonic mitochondria are inherited maternally and independently of the nuclear genome. They show limited activity during the early developmental stages before embryonic genome activation. It has been shown that in vitro culture (IVC) has an adverse effect on mitochondrial function in embryos. So far several attempts have been performed to improve and rescue the impaired mitochondria. It has been shown that vitamin K2 (a membrane-bound electron carrier, similar to ubiquinone) was used to rescue mitochondrial dysfunction and resulted in more efficient ATP production in eukaryotic cells (Vos et al. 2012 Science 336, 1306–1310). Therefore, the aim of the present study was to investigate the effects of supplementation of vitamin K2 on mitochondrial activity and blastocyst rate. Cumulus–oocytes complexes (n = 687) recovered from slaughtered animals, were matured and fertilized in vitro according to our standard procedures. After fertilization, zygotes were cultured in SOF media supplemented with 10 mg mL–1 BSA. At 96 h post-fertilization, vitamin K2 was added to the culture media (n = 448 oocytes). On Day 7, treatment embryos were compared with untreated controls (n = 239 oocytes). In vitro culture was carried out at 38.5°C under 5% CO2, 7% O2, and 88% N2. Differences among groups in blastocyst yield were analysed by ANOVA. Mitochondrial activity data was analysed by unpaired 2-tailed t-tests. Results show that the vitamin K2-treated group had a significantly (P < 0.05) higher blastocyst rate (+8.6%), expanded blastocyst rate (+7.8%), as well as better morphological quality compared with the control group. Furthermore, to evaluate mitochondria activity, pools of embryos of each treatment were labelled with a specific dye for active mitochondria (Mitotracker Red). A significantly higher intensity of Mitotracker Red (P < 0.05) was observed in the vitamin K2 treatment versus control group, as measured by fluorescent microscopy. In conclusion, for the first time, our data prove that supplementation of vitamin K2 during IVC of bovine embryos increases blastocyst rates and embryo quality. Future studies will focus on gene expression to identify targets implicated in impaired mitochondrial activity in in vitro bovine embryo production.

75 Improvement of Developmental Competence of Bovine In Vitro-Produced Embryos by Using Charcoal:Dextran-Stripped Fetal Bovine Serum on In Vitro Culture Media

2018 ◽  
Vol 30 (1) ◽  
pp. 176
Author(s):  
A. Mesalam ◽  
R. Kong ◽  
B.-H. Choi ◽  
K.-L. Lee ◽  
B.-Y. Park ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Developmental Competence ◽  
Mitochondrial Activity ◽  
Mrna Levels ◽  
Fetal Bovine

Serum has widely been used as a main supplement to embryo in vitro culture media as it contains embryotrophic factors. Charcoal:dextran treatment of fetal bovine serum (FBS) removes lipophilic chemicals and certain steroid hormones and growth factors. The objective of this study was to investigate the effects of charcoal:dextran-stripped fetal bovine serum (CDS FBS) and heat-inactivated FBS (HI FBS) in embryo culture medium (SOF-BE1 medium supplemented with 10% of serum) on their ability to support in vitro development of bovine embryos. The developmental ability and quality of bovine embryos were determined by assessing their cell number, lipid content, mitochondrial activity, gene expression, and cryo-tolerance. The experiment was conducted in 6 replicates (350 oocytes per group). The differences in embryo development, integrated optical intensity, and expression levels of the various genes between experimental groups were analysed by one-way ANOVA. Duncan’s multiple range tests were used to test the differences between the treatments. The level of statistical significance was set at P < 0.05. The percentages of embryos that underwent cleavage and formed a blastocyst were significantly (P < 0.05) higher in medium containing CDS FBS than in medium containing HI FBS (42.84 ± 0.78% v. 36.85 ± 0.89%, respectively). The total number of cells per Day 8 blastocyst was not significantly different (P > 0.05) between the CDS FBS group (208.40 ± 14.77) and the HI FBS group (195.11 ± 19.15). Furthermore, the beneficial effects of CDS FBS on embryos were associated with a significantly increased mitochondrial activity, as identified by MitoTracker Green, and reduced intracellular lipid content, as identified by Nile red staining, which increased their cryo-tolerance. The post-thaw survival rate of blastocysts was significantly (P < 0.05) higher after 24 h in the CDS FBS than in the HI FBS group (85.33 ± 4.84% v. 68.67 ± 1.20%). Quantitative reverse transcription PCR showed that the mRNA levels of lipid metabolism-related genes, acyl-CoA synthetase long-chain family member 3, acyl-coenzyme A dehydrogenase long-chain, and the cholesterol metabolism related gene hydroxymethylglutaryl-CoA reductase were significantly increased upon culture with CDS FBS. Moreover, the mRNA levels of survival gene sirtuin 1, antioxidant gene superoxide dismutase 2, and anti-apoptotic associated gene B-cell lymphoma 2 in frozen–thawed blastocysts were significantly (P < 0.05) higher in the CDS FBS group than in the HI FBS group; however, the mRNA level of the pro-apoptotic gene BCL2-associated X protein was significantly reduced. In conclusion, these data suggest that supplementation of in vitro culture medium with CDS FBS improves in vitro bovine embryo developmental competence and the quality of blastocysts in terms of their crytolerance and gene expression. This research was supported by grant from the Next-Generation BiogGeen21 (No. PJ01107703), IPET (No. 315017-5 and 117029-3), Allergy free cat (Co.. Felix Pets), BK21plus, and KGSP.

A novel method for toxicology: In vitro culture system of a rat preantral follicle

2011 ◽  
Vol 27 (7) ◽  
pp. 637-645 ◽  
Author(s):  
Wan Xuying ◽  
Zhu Jiangbo ◽  
Zhu Yuping ◽  
MA Xili ◽  
Liu Zhen ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture System ◽  
Preantral Follicle ◽  
In Vitro Culture System ◽  
Novel Method

scholarly journals The Effect of Penicillin on the Vitality of Bull Spermatozoa

2020 ◽  
Vol 2 (3) ◽  
pp. 30-34
Author(s):  
Ján Kováč ◽  
Nishonoy Khasanova ◽  
Tomáš Slanina ◽  
Peter Massányi ◽  
Eva Tvrdá
Keyword(s):  
In Vitro Culture ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Lower Amount ◽  
Mitochondrial Activity ◽  
Sperm Analysis ◽  
Sperm Motion ◽  
Acrosomal Integrity

Antibiotic supplementation into semen extenders is an important way to control several microorganisms that can affect semen quality by their presence. The objective of the present work is to estimate the effect of two different concentrations (300 µg/mL and 600 µg/mL) of penicillin on the selected quality parameters of spermatozoa collected from bulls (motility, mitochondrial activity, acrosome integrity and membrane integrity) after 0, 2 and 24 h of in vitro culture. Sperm motion was examined using HTM IVOS computer-aided sperm analysis (CASA), cell viability was assessed with the metabolic activity (MTT) assay. The acrosomal integrity was evaluated following the fast green – rose bengal staining protocol and the eosin – nigrosin staining method was used to assess the functional integrity of the sperm membrane. Our results indicate that penicillin at lower amount significantly (p>0.05) decreased the sperm motility, mitochondrial activity and membrane integrity after 24 h of in vitro culture. Supplementation of higher doses of this substance led to a significant decrease of the sperm motion during 0, 2 (p>0.05) as well as after 24 h (p>0.01), of the viability after 2 h (p>0.05) and 24 h (p>0.01), of the acrosomal integrity after 24 h (p>0.05) and of the membrane integrity at 24 h (p>0.001) too. We can consider, that the effect of penicillin addition to bovine spermatozoa during in vitro incubation is time and dose dependent.  

Growth differentiation factor‐9 improves development, mitochondrial activity and meiotic resumption of sheep oocytes after in vitro culture of secondary follicles

2019 ◽  
Vol 54 (9) ◽  
pp. 1169-1176 ◽  
Author(s):  
Alane P. O. Monte ◽  
Jamile M. Santos ◽  
Vanúzia G. Menezes ◽  
Bruna B. Gouveia ◽  
Thae L. B. G. Lins ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Mitochondrial Activity ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9

scholarly journals l-Carnitine Supplementation during In Vitro Maturation and In Vitro Culture Does not Affect the Survival Rates after Vitrification and Warming but Alters Inf-T and ptgs2 Gene Expression

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
Keyword(s):  
Gene Expression ◽  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Survival Rates ◽  
Cell Number ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Bovine Embryo ◽  
Carnitine Supplementation

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.

In vivo and in vitro strategies to support caprine preantral follicle development after ovarian tissue vitrification

2018 ◽  
Vol 30 (8) ◽  
pp. 1055 ◽  
Author(s):  
N. J. Donfack ◽  
K. A. Alves ◽  
B. G. Alves ◽  
R. M. P. Rocha ◽  
J. B. Bruno ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Preantral Follicle ◽  
Hormone Production ◽  
Treatment Groups ◽  
Morphology Development ◽  
Fresh Culture

The aim of the present study was to compare fresh and vitrified goat ovarian tissue after autotransplantation and in vitro culture. Adult goats were completely ovariectomised and each ovarian pair was sliced and distributed among six different treatment groups: fresh control, fresh transplant, fresh culture, vitrified control, vitrified transplant and vitrified culture. Follicular morphology, development, growth, density, revascularisation and hormone production were evaluated in all groups. Three antral follicles (two in the fresh transplant and one in the vitrified transplant groups) were observed on the surface of the graft 90 days after transplantation. The percentage of morphologically normal follicles was similar in the fresh control, fresh transplant and vitrified transplant groups. The percentage of developing (transition, primary and secondary) follicles was higher after in vitro culture of fresh or vitrified tissue. Transplantation resulted in a lower follicle density. Serum oestradiol concentrations remained constant during the entire transplantation period. In contrast, progesterone production decreased significantly. Expression of CD31 mRNA was lower in fresh culture. In conclusion, restoration of goat ovarian function can be successfully achieved following transplantation of both fresh and vitrified goat ovarian tissue. However, transplantation induced higher follicle loss than in vitro culture.

scholarly journals Control of growth and development of preantral follicle: insights from in vitro culture

2018 ◽  
Vol 15 (Suppl. 1) ◽  
pp. 648-659
Author(s):  
José Ricardo de Figueiredo ◽  
Laritza Ferreira de Lima ◽  
José Roberto Viana Silva ◽  
Regiane Rodrigues Santos
Keyword(s):  
In Vitro Culture ◽  
Growth And Development ◽  
Preantral Follicle
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