Porcine oocyte maturation in vitro under different conditions

2006 ◽  
Vol 3 (2) ◽  
pp. 95-100
Author(s):  
Hu Jun-He ◽  
Yang Chun-Rong ◽  
Dou Zhong-Ying
Keyword(s):  
Oocyte Maturation ◽  
Culture Medium ◽  
Follicle Stimulating Hormone ◽  
Human Chorionic Gonadotrophin ◽  
Serum Free Medium ◽  
Free Medium ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Pregnant Mare Serum Gonadotrophin

AbstractThe effects of hormone additions at various stages and different basic media, with or without serum, on porcine oocyte maturation in vitro were studied. The results showed that the rate of maturation was not significantly different with three different stages of hormone supplement; the rate of maturation on modified TCM199 medium (54.01%) was higher than that on TCM199 (46.16%) and (47.14%), but these differences were not significant; and the rate of maturation on serum-free medium (67.10%) was significantly higher than that on medium plus serum (52.22%). Therefore, modifed tissue culture medium 199 (mTCM199)+10I U/ml pregnant mare serum gonadotrophin (PMSG)+10I U/ml human chorionic gonadotrophin (hCG)+2.5 IU/ml follicle stimulating hormone (FSH) was a suitable medium for culture of porcine oocytes in vitro, and the rate of maturation was 67.10%.

173 MELATONIN ALLEVIATES THE ENDOPLASMIC RETICULUM STRESS THROUGH THE REGULATING OF UNFOLDING PROTEIN RESPONSE SIGNALING DURING PORCINE OOCYTE MATURATION IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 195 ◽  
Author(s):  
J.-Y. Park ◽  
H.-J. Park ◽  
J.-W. Kim ◽  
S.-Y. Park ◽  
S.-G. Yang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Er Stress ◽  
Oocyte Maturation ◽  
Caspase 3 ◽  
Oxygen Species ◽  
Melatonin Treatment ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Protein Response

Unfolding protein response (UPR) is a defence mechanism during endoplasmic reticulum (ER) stress in mammalian cells. Especially, UPR genes and regulation of reactive oxygen species is involved in ER stress response on porcine oocyte maturation in vitro. Some studies have shown that melatonin treatment results in reducing oxidative stress, a protective function of free radical damage in oocyte maturation and embryo development. Also, melatonin has an important role in reducing reactive oxygen species and ER stress. However, it is unknown how the changes of UPR genes expression levels are affected the porcine oocyte maturation. In addition, there are no reports about ER stress recovery mechanism by melatonin during porcine oocyte maturation. Here, we investigated the UPR signal genes (Bip/Grp78, Atf4, p90/p50Atf6, and Xbp1) and ER-stress mediated apoptosis factors (Chop and Cleaved caspase 3) in porcine oocyte maturation in vitro. Expression of Chop and Cleaved caspase 3 mRNA levels were significantly increased (P < 0.01) in matured oocytes (metaphase II; 44 h) in vitro. Porcine oocytes were cultured in maturation medium with ER stress inducer, tunicamycin (Tm), and supplemented with various concentrations (1, 5, and 10 μg mL−1) of Tm for 0 to 44 h. Our results indicated that the proportion of matured oocytes was significantly decreased in Tm-treated groups in a dose-dependent manner (60.1 ± 1.3, 46.5 ± 2.1, and 38.9 ± 5.1% at 1, 5, and 10 μg mL−1 of Tm) compared with the control group (76.6 ± 1.9%). Likewise, mRNA expression of UPR regulator genes (Grp78/Bip, Aft4, Xbp1, Chop, and Cleaved caspase 3) was decreased by melatonin treatment (0.1 μM, 22–44 h) after pretreatment of Tm (5 μg mL−1, 0–22 h) during oocyte maturation. Our results demonstrated that the roles of melatonin as UPR signaling regulator for reducing ER stress are essential for promotion of porcine oocyte maturation and cumulus cell expansion of cumulus-oocyte complex. Moreover, the current study was initiated to confirm a functional link between effect of melatonin and regulating of UPR signaling in porcine oocytes maturation. These results suggest that melatonin improve the oocyte maturation and cumulus cells expansion by regulating of UPR signal genes against the ER stress during the porcine in vitro maturation process. This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01117604) and the Bio-industry Technology Development Program (316037–04–1-HD020) through the Rural Development Administration, the Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

Fasciola hepatica in vitro: increased susceptibility to fasciolicides in a defined serum-free medium

Parasitology ◽  
1987 ◽  
Vol 95 (1) ◽  
pp. 165-171 ◽  
Author(s):  
D. C. Jenkins ◽  
P. Topley ◽  
E. B. Rapson
Keyword(s):  
Culture Medium ◽  
Fasciola Hepatica ◽  
Liver Microsomes ◽  
Metabolic Activation ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Highly Active ◽  
Mammalian Liver

SUMMARYThe cidal properties of some phenolic, halogenated diphenyl, salicylanilide, benzimidazole and diaminophenoxyalkane anthelmintics, against 6-week-old worms of Fasciola hepatica were assessed in vitro. In a conventional fluke culture medium containing RPMI 1640, supplemented with serum with or without rabbit erythrocytes or pink-ghosts, only the halogenated diphenyl and salicylanilide compounds showed activity at concentrations equal to or less than 100 μm. However, when basal, serum and cell-free RPMI 1640 was used, all compounds other than diamphenethide were highly active, their minimum lethal concentrations being some 25–125 times lower under these conditions. The inclusion of rabbit liver microsomes in the basal culture medium resulted in diamphenethide exhibiting cidal activity equivalent to that seen when its free-amine active metabolite was assayed. The possibility that the activity of many of these compounds was masked in vitro because of their serum binding properties is discussed. Recommendations are made that in vitro screens for new fasciolicides should be carried out in serum-free medium and that additional replicates containing mammalian liver microsomes and liver cytosolic extracts be included as means for the metabolic activation of certain otherwise undetectable prodrugs.

scholarly journals Elevation of MPF and MAPK gene expression, GSH content and mitochondrial distribution quality induced by melatonin promotes porcine oocyte maturation and development in vitro

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9913
Author(s):  
Zimo Zhao ◽  
Ling Yang ◽  
Dan Zhang ◽  
Zi Zheng ◽  
Ning Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Developmental Potential ◽  
Expression Levels ◽  
Maturation In Vitro ◽  
Porcine Oocyte ◽  
Maturation Rate ◽  
Mitochondrial Distribution ◽  
Mapk Gene

The MPF and MAPK genes play crucial roles during oocyte maturation processes. However, the pattern of MPF and MAPK gene expression induced by melatonin (MT) and its correlation to oocyte maturation quality during the process of porcine oocyte maturation in vitro remains unexplored. To unravel it, in this study, we cultured the porcine oocytes in maturation medium supplemented with 0, 10−6, 10−9, and 10−12 mol/L melatonin. Later, we analyzed the MPF and MAPK gene expression levels by RT-PCR and determined the maturation index (survival and maturation rate of oocytes). The GSH content in the single oocyte, and cytoplasmic mitochondrial maturation distribution after porcine oocyte maturation in vitro was also evaluated. We also assessed the effects of these changes on parthenogenetic embryonic developmental potential. The oocytes cultured with 10−9mol/L melatonin concentration showed higher oocyte maturation rate, and MPF and MAPK genes expression levels along with better mitochondrial distribution than the 0, 10−6, and 10−12 mol/L melatonin concentrations (p < 0.05). No significant difference was observed in the survival rates when the oocytes were cultured with different melatonin concentrations. The expression of the MPF gene in the oocytes cultured with 10−6 mol/L melatonin was higher than with 10−12 and 0 mol/L melatonin, and the expression of the MAPK gene in 10−6 and 10−12 group was higher than the control (p < 0.05). As far as the embryonic developmental potential is concerned, the cleavage and blastocyst rate of oocytes cultured with 10−6 and 10−9 mol/L melatonin was significantly higher than the 10−12 mol/L melatonin and control. In conclusion, 10−9–10−6 mol/L melatonin significantly induced the MPF and MAPK gene expression; besides, it could also be correlated with GSH content of single oocyte, mitochondrial maturation distribution, and the first polar body expulsion. These changes were also found to be associated with parthenogenetic embryo developmental potential in vitro.

172 THE APOPTOTIC EFFECTS OF BISPHENOL A-INDUCED MITOCHONDRIAL-DERIVED REACTIVE OXYGEN SPECIES ON MATURATION OF PORCINE OOCYTES IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 194
Author(s):  
S.-Y. Park ◽  
H.-J. Park ◽  
J.-W. Kim ◽  
J.-Y. Park ◽  
S.-G. Yang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Critical Role ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Meiotic Maturation ◽  
Control Group ◽  
Ros Production ◽  
Maturation In Vitro ◽  
Porcine Oocyte

Bisphenol A (BPA) is well known as oestrogen-like chemical and it is widely used in plastic products. Many studies have reported that BPA exposure has a well-known toxicity effect on reproduction function, such as reducing the number of ovulated oocytes, oocyte quality, and maturation rate. Recently, BPA induced mitochondrial-derived reactive oxygen species (mito-ROS) and disrupted mitochondrial homeostasis by increasing of superoxide anions production. In this study, we investigated how the regulation of mito-ROS production may play a critical role in meiotic maturation and expansion of cumulus cells during the in vitro maturation progression of porcine oocytes. Furthermore, we investigated the toxicity effect of BPA exposure on mitochondrial functions and mito-ROS production during porcine oocyte maturation in vitro. All results were analysed using a 1-way ANOVA followed by Bonferroni’s and Tukey’s Multiple Comparison Test and t-tests. First, porcine oocytes were matured in NCSU-23 medium supplemented with BPA (50, 75, and 100 µM) for 44 h. Our results indicated that the rates of matured oocytes were significantly decreased by BPA exposure in a dose-dependent manner (69.4 ± 5.1, 50.9 ± 6.3, and 29.9 ± 5.8% for BPA treatments of 50, 75, and 100 μM) compared with control group (70.2 ± 7.8%; P < 0.05). Next, we confirmed the secretion functions of oocyte and cumulus cell of cumulus-oocyte complex (COC) and ROS production. Cumulus cell secretion factors (has2, tnfaip6, and cx37) mRNA expression in COC were decreased in the BPA-treated (75 µM) group. In addition, mRNA expressions of mitochondrial-specific antioxidant enzymes (sod2, P < 0.001; prdx3, P < 0.01; prdx5, P < 0.001) and mitochondrial apoptosis genes (bax and caspase-3, P < 0.01) were significantly increased in COC of the BPA-treated (75 µM) group. We measured mitochondrial membrane potential and mito-ROS production using JC-1 analysis and Mito-SOX staining, respectively. The BPA treatment caused a rapid decrease of mitochondrial membrane potential maintenance and increase of mito-ROS production in porcine COC. Moreover, mitochondrial-specific ROS scavenger, Mito-Tempo (0.1 µM) treatment was significantly increased the meiotic maturation of porcine oocytes compared with control group (78.5 ± 3.5 v. 65.8 ± 5.0%; P < 0.05). Based on these results, we first confirmed that BPA exposure reduces the meiotic maturation and cumulus cells expansion of COC by increasing mito-ROS production during porcine oocyte maturation in vitro. Therefore, controlling of mito-ROS for mitochondrial function maintenance and apoptosis plays a critical role in improving porcine oocyte maturation in vitro. This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01117604) and the Bio-industry Technology Development Program (316037–04–1-HD020) through the Rural Development Administration, the Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

Bovine blastocyst development after follicle-stimulating hormone and platelet-derived growth factor treatment for oocyte maturation in vitro

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 27-34 ◽  
Author(s):  
Kathleen M. Harper ◽  
Benjamin G. Brackett
Keyword(s):  
Growth Factor ◽  
Oocyte Maturation ◽  
Follicle Stimulating Hormone ◽  
Positive Influence ◽  
Platelet Derived Growth Factor ◽  
Blastocyst Development ◽  
Maturation In Vitro ◽  
Igf I ◽  
Stimulating Hormone

SummaryBovine embryonic viability after in vitro fertilisation (IVF) was affected by addition of platelet-derived growth factor (PDGF) or growth factor combinations to gonadotrophin-supplemented serum-free medium for in vitro maturation (IVM). Thus, 0.1, 1.0 or 10.0 ng PDGF/ml combined with 500 ng follicle-stimulating hormone (FSH)/ml enhanced oocyte maturation as reflected by greater proportions (p < 0.05) of matured and inseminated oocytes developing to blastocysts in vitro than were afforded by PDGF alone, i.e. 32.4%, 28.2% and 31.0% respectively vs. 11.4%, 13.5% and 8.6% respectively. By contrast, luteinising hormone (LH, 500 ng/ml) combined with the same concentrations of PDGF did not increase the proportions of oocytes developing to blastocysts (16.2%, 12.5% and 16.9%). Epidermal growth factor (EGF) + insulin-like growth factor-I (IGF-I) with or without PDGF when added to low concentrations of FSH or LH did not significantly improve cleavage or morula development. Significant improvement (p & 0.05) in blastocyst formation was seen when FSH was supplemented with either EGF (38.0%) or EGF + IGF-I + PDGF (40.7%) compared with FSH alone (27.4%) or FSH + LH (24.3%), but no improvement followed FSH + EGF + IGF-I (33.6%) or FSH + LH + EGF + IGF-I + PDGF (29.2%) treatments for IVM. Results revealed a positive influence on blastocyst development of FSH and PDGF or EGF ± PDGF during IVM and suggest the possibility of a concerted action of gonadotrophins with growth factors in physiological (functional) oocyte maturation.

Toxic effects and possible mechanisms of hydrogen sulfide and/or ammonia on porcine oocyte maturation in vitro

2018 ◽  
Vol 285 ◽  
pp. 20-26 ◽  
Author(s):  
Lei-Lei Yang ◽  
Yong Zhao ◽  
Shi-Ming Luo ◽  
Jun-Yu Ma ◽  
Zhao-Jia Ge ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Oocyte Maturation ◽  
Toxic Effects ◽  
Maturation In Vitro ◽  
Porcine Oocyte

GDF8 activates p38 MAPK signaling during porcine oocyte maturation in vitro

2017 ◽  
Vol 101 ◽  
pp. 123-134 ◽  
Author(s):  
Junchul David Yoon ◽  
Seon-Ung Hwang ◽  
Eunhye Kim ◽  
Minghui Jin ◽  
Soochong Kim ◽  
...  
Keyword(s):  
P38 Mapk ◽  
Oocyte Maturation ◽  
Mapk Signaling ◽  
Maturation In Vitro ◽  
Porcine Oocyte

scholarly journals The Effects of Polyadenylation Status on MPFs During In Vitro Porcine Oocyte Maturation

2016 ◽  
Vol 39 (5) ◽  
pp. 1735-1745 ◽  
Author(s):  
Huiyu Liu ◽  
Yan Gao ◽  
Bo Zhai ◽  
Hao Jiang ◽  
Yu Ding ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Cyclin B1 ◽  
Spindle Formation ◽  
Large Follicle ◽  
Maturation In Vitro ◽  
First Polar Body ◽  
Chromosome Alignment ◽  
Porcine Oocyte ◽  
Maternal Mrnas

Aims: This study aims to clarify the effects of polyadenylation status on M-phase promoting factors (MPFs) during in vitro porcine oocyte maturation. Methods: In this study, porcine follicular oocytes from large follicles (> 5 millimeter (mm)) and small follicles (< 3 mm) were examined at different follicular developmental stages. The polyadenylation of maternal mRNAs was inhibited by the addition of 3'-deoxyadenosine (3'-da) during the germinal vesicle (GV)(0 h), GV breakdown (GVBD)(18 h), metaphase I (MI)(28 h), and metaphase II (MII) (44 h) stages. In addition, the expression levels and poly-(A) tail lengths of the maternal mRNAs Cyclin B1 and cell division cycle 2 (Cdc2) were determined by real-time quantitative PCR. Immunofluorescence was used to assess spindle formation and chromosome alignment in the examined oocytes. Results: In large-follicle oocytes, the effects of inhibiting polyadenylation caused the percentage of mature to be significantly lower for the treated group than for the untreated group (p < 0.01). 3'-da can significantly improve the rate of small oocyte maturation in vitro and inhibits Cdc2 polyadenylation. Cyclin B1 plays a significant role in promoting the maturation of large-follicle oocytes. Polyadenylation contributes to the formation of dominant follicles and facilitates the selection of dominant follicles. However, the inhibition of adenylation affected spindle formation-related propulsion and chromosome alignment in both large- and small-follicle oocytes. The first polar body could not be extruded in certain large follicles. Conclusions: 3'-da can significantly improve the rate of small oocyte maturation in vitro, but it can also affect spindle formation-related propulsion and chromosome alignment.

scholarly journals THE REQUIREMENT FOR HYDROCORTISONE IN ANTIBODY-FORMING TISSUE CULTIVATED IN SERUM-FREE MEDIUM

1964 ◽  
Vol 119 (6) ◽  
pp. 1027-1049 ◽  
Author(s):  
Charles T. Ambrose
Keyword(s):  
Culture Medium ◽  
Essential Amino Acids ◽  
Secondary Response ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Secondary Antibody ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

It was previously reported from this laboratory that the secondary antibody response can regularly be elicited in vitro from fragments of rabbit lymph node node cultured in Eagle's medium supplemented with normal rabbit serum. Evidence is now presented that physiological levels of hydrocortisone (0.01 to 1.0 µM) can substitute for serum in the culture medium. However, with the omission of serum, serine (0.1 mM) must be included among Eagle's "essential" amino acids for consistent optimal antibody production. In some experiments the addition of insulin (0.5 unit/ml) and vitamin B12 (0.5 µg/ml) has further enhanced the secondary response in this serum-free medium.

Specificity of inhibition by steroids of porcine oocyte maturation in vitro

1979 ◽  
Vol 209 (1) ◽  
pp. 81-90 ◽  
Author(s):  
Joel D. Richter ◽  
Robert W. McGaughey
Keyword(s):  
Oocyte Maturation ◽  
Maturation In Vitro ◽  
Porcine Oocyte
Sign in / Sign up

Export Citation Format

Share Document