287 EFFECT OF CYSTEAMINE DURING IN VITRO MATURATION OF OPU DERIVED BOVINE OOCYTES ON FURTHER IN VITRO EMBRYONIC DEVELOPMENT AND PREGNANCY RATE

2006 ◽  
Vol 18 (2) ◽  
pp. 251
Author(s):  
J. S. Merton ◽  
B. Landman ◽  
E. Mullaart
Keyword(s):  
Embryonic Development ◽  
Pregnancy Rate ◽  
Production Rate ◽  
In Vitro Maturation ◽  
Radical Scavenging ◽  
Protective Role ◽  
Embryo Production ◽  
Bovine Oocytes ◽  
In Vitro Embryonic Development

Glutathione (GSH) plays an important protective role in relation to reactive oxygen species generated by normal oxidative metabolism in the cell. The presence of cysteamine during in vitro maturation may facilitate the synthesis of GSH by immature oocytes. In a previous study we showed a positive effect of the presence of cysteamine during in vitro maturation of slaughterhouse derived bovine oocytes on subsequent in vitro embryonic development (Merton et al. 2004 Rep. Fert. Dev. 16, 279 abstract). This report shows the results of a field trial with ultrasound guided transvaginal oocyte collection (OPU) derived oocytes, in order to confirm our previous results obtained with slaughterhouse derived oocytes. Immature cumulus–oocyte complexes (COCs) were recovered twice weekly by ovum pick-up (OPU) at two collection centres from 11 cows and 147 pregnant heifers. COCs were matured in vitro in TCM199/FCS/LH/FSH supplemented either with or without cysteamine (0.1 mM). Subsequently, matured oocytes were fertilised with frozen-thawed gradient-separated semen and further cultured for 7 days in SOFaaBSA. At Day 7, Morula grade 1 (IETS) were transferred fresh and early-, mid- and exp-Blast grade 1 and 2 were transferred either fresh or frozen/thawed. The experimental design was a 2 × 2 factorial. Results were analysed by Chi-square analyses. The results show that the presence of cysteamine during in vitro maturation significantly affected embryo production from OPU derived COCs (23.4% and 34.4% Morula + Blastocyst rate at Day 7 for control and cysteamine, respectively; Table 1). This higher embryo production rate was mainly due to an increased number of Blastocysts. Also the proportion of grade 3 embryos was significantly reduced in the cysteamine group (P < 0.01). The number of transferable embryos per session was 1.06 and 1.73 for control and cysteamine, respectively. Pregnancy rate was not significantly affected by the presence of cysteamine during in vitro maturation for both fresh and frozen/thawed embryos (fresh: 40.5% and 44.8%, frozen/thawed: 44.4% and 47.2% for control and cysteamine, respectively). These results show that the presence of cysteamine during in vitro maturation affects further in vitro embryonic development, resulting in a higher embryo production rate. This suggest that an apparently ‘simple’ extra protection of the oocyte, due to the free radical scavenging potency of GSH, can have an enormous effect (63.2% relative increase in transferable embryos) on its in vitro developmental potency. The intrinsic quality of the ‘extra’ produced transferable embryos seems not to be different, since pregnancy rate was not affected. Table 1. Effect of cysteamine during in vitro maturation of OPU-derived bovine oocytes on subsequent in vitro embryonic development

scholarly journals 320EFFECT OF CYSTEAMINE DURING IN VITRO MATURATION ON FURTHER EMBRYONIC DEVELOPMENT AND POSTTHAW SURVIVAL OF IVP BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 279
Author(s):  
J.S. Merton ◽  
M. Gerritsen ◽  
D. Langenbarg ◽  
Z.L. Vermeulen ◽  
T. Otter ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Production Rate ◽  
In Vitro Maturation ◽  
Survival Rates ◽  
Protective Role ◽  
Bovine Embryos ◽  
Embryo Production ◽  
In Vitro Embryonic Development ◽  
In Vitro Survival

The uptake of cysteamine by immature oocytes may facilitate the synthesis of glutathione (GSH) during in vitro maturation, as reported by Matos et al. (1995 Mol. Reprod. Dev. 42 432–436). GSH plays an important protective role in relation to reactive oxygen species generated by normal oxidative metabolism. This study investigated the effects of the presence of cysteamine during in vitro maturation on subsequent in vitro embryonic development and postthaw in vitro survival. Immature Cumulus-Oocyte-Complexes (COCs) were recovered from ovaries 6 to 8h after slaughter. COCs were matured in vitro for 22 to 24h in TCM199/FCS/LH/FSH supplemented either with or without cysteamine (0.1mM), Subsequently, matured oocytes were fertilized with frozen-thawed Percoll-separated semen and further cultured for seven days in SOFaaBSA. Morulae grade 1 (IETS) and blastocysts grades 1 and 2 (IETS) were frozen on Day 7 in 10% Glycerol using a conventional slow freezing procedure (Wagtendonk-de Leeuw et al. 1995 Cryobiology;; 32 157–167). In vitro survival was measured by rates of blastocyst formation and reexpansion at 24h and hatching/ed blastocysts at 72h in SOFaaBSA supplemented with 5% FCS. Results were analyzed by Chi-square analyses. The presence of cysteamine during in vitro maturation significantly affected the embryo production rate (19.4% and 24.0% for control and cysteamine at Day 7, respectively). The higher number of embryos at Day 7 was totally due to an increased number of blastocysts (Table 1); however, the distribution of embryos among the different quality grades was not affected. Addition of cysteamine did not affect the post thaw survival of the frozen/thawed embryos (85% v. 91% reexpansion and 33% v. 34% hatching/ed for control v. cysteamine, respectively). These results show that the presence of cysteamine during in vitro maturation, does affect further in vitro embryonic development, resulting in a higher embryo production rate. Embryo quality, expressed in morphological grades and postthaw survival rates, were not affected. A field trial will be conducted in order to confirm these results with ovum pick up-derived oocytes. Table 1 Effect of cysteamine during in vitro maturation on subsequent in vitro embryonic development of IVP bovine embryos (number of replicates: 5)

137 Kinetics of In Vitro Embryonic Development According to the Follicular Population in Bos Indicus

2018 ◽  
Vol 30 (1) ◽  
pp. 208
Author(s):  
J. G. Soares ◽  
F. M. Morato ◽  
G. F. Rossi ◽  
B. M. Bayeux ◽  
A. S. Oliveira ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Formation Rate ◽  
Bos Indicus ◽  
Prostaglandin F2α ◽  
Lower Percentage ◽  
Embryo Production ◽  
High Category ◽  
Kinetics Of ◽  
In Vitro Embryonic Development

The present study aimed to evaluate the effect of the follicular population from cull Nelore (Bos indicus) on the kinetics of in vitro embryonic development. At random stages of the oestrous cycle (Day –5), a total of 28 cull Nelore cows were synchronized with an intravaginal progesterone device associated with oestradiol benzoate (2.0 mg IM). At the same moment, a dose of prostaglandin F2α (2.0 mg im) was also administered to promote luteolysis and absence of corpus luteum (CL) at the time of ovum pick-up (OPU). Five days later (Day 0), all cows underwent an OPU session and the recovered oocytes were submitted to in vitro embryo production (IVEP). The same procedures were repeated 2 times at 30-day intervals (Day 25 and Day 55). Semen from a single batch of a previously tested bull was used for all IVEP. Blastocyst production and hatching were verified on Days 7, 8, and 9 of the IVEP. Data were analysed by the GLIMMIX procedure of SAS 9.3® (SAS Institute Inc., Cary, NC, USA). Data of the 3 OPU sessions were grouped and the cows were classified into 3 categories according to the follicular population: Low (19.7 ± 0.9 follicles, n = 27), Medium (33.5 ± 0.8 follicles, n = 29), and High (58.7 ± 3.2 follicles, n = 28). The Low category had a lower rate of viable oocytes [(number of viable oocytes/total number of oocytes) × 100; 62.0 v. 69.5%; P = 0.02] and cleavage rate [(number of cleaved/total number of oocytes) × 100; 55.9 v. 66.8%; P = 0.001] than the High category. The blastocyst formation rate [number of blastocysts/total number of oocytes) × 100] on Day 7 and Day 8 was lower for Low and Medium compared with the High category (Day 7: 26.1b v. 29.0b v. 35.1a %; P = 0.001; and Day 8: 29.2b v. 30.2b v. 34.7a %; P = 0.05). No differences were found in blastocysts rate on Day 9 among Low, Medium, and High categories (14.1 v. 15.9 v. 16.2%; P = 0.61). However, Low category had a lower percentage of hatched blastocysts [(number of hatched blastocysts/number of blastocysts) × 100] on Day 7 compared with High category (2.9 v. 12.0%; P = 0.01). These results reported that cull Nelore with High follicular population showed higher rates of embryo production and hatched blastocysts compared with cows with Low follicular population. We concluded that the kinetics of in vitro embryonic development was compromised in cull Nelore (Bos indicus) with low follicular population submitted to OPU-IVEP. This research was supported by Fapesp 2012/50533–2 (GIFT).

In Vitro Maturation of Bovine Oocytes with Bovine Herpesvirus 5 Does Not Compromise the Subsequent Embryonic Development.

2009 ◽  
Vol 81 (Suppl_1) ◽  
pp. 227-227
Author(s):  
Camila Silva ◽  
Alicio Martins ◽  
Renata Sanches Calegari ◽  
Marcelo Carnelli Frade ◽  
Diego Gouvea Souza ◽  
...  
Keyword(s):  
Embryonic Development ◽  
In Vitro Maturation ◽  
Bovine Herpesvirus ◽  
Bovine Oocytes

scholarly journals IN VITRO MATURATION OF BOVINE OOCYTES FOR IN VITRO FERTILIZATION

2009 ◽  
Vol 15 (1) ◽  
pp. 16-19
Author(s):  
Ioan GROZA ◽  
Simona CIUPE ◽  
Mihai CENARIU ◽  
Emoke PALL ◽  
Anamaria PETREAN
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
In Vitro Maturation ◽  
Cultivation Conditions ◽  
Vitro Fertilization ◽  
Morphological Evaluation ◽  
17Β Estradiol ◽  
Bovine Oocytes

The objective of the present study was to asses the quality of various cultivation media used for the maturation of bovine oocytes that are prepared for IVF. Upon collection from slaughtered bovine ovaries and after morphological evaluation, a total number of 513 viable oocytes have been selected for cultivation, being divided into 3 batches, 171 oocytes / batch. The oocytes belonging to batch 1 were cultivated in TCM 199 NaHCO3 + 10% FCS + FSH 20 μl/ml. The oocytes belonging to batch 2 were cultivated in TCM 199 NaHCO3 + 10% FCS + HCG 2.3 x 103 UI/ml + FSH 8 μl/ml + pyruvate 0.25 mM + 17β estradiol 1 μl/ml. The oocytes belonging to batch 3 were cultivated in TCM 199 NaHCO3 + 10% FCS + 17β estradiol 1 μl/ml + FSH 20 μl/ml. The cultivation conditions, for all three batches, were: 24 hours at 39°C, 5% CO2. Spermatozoa have been prepared using the Percoll method and IVF of the matured oocytes has been performed. Embryonic development has been assessed 72 hours and then up to 10 days after IVF. The results showed the superior quality of the oocytes belonging to batch 2 and matured using TCM 199 NaHCO3 + 10% FCS + HCG 2.3x103 UI/ml + FSH 8 μl/ml + pyruvate 0.25 mM + 17β estradiol 1 μl/ml, as their use for IVF yielded the highest number of viable embryos.

Effects of supplementation of medium with different antioxidants during in vitro maturation of bovine oocytes on subsequent embryo production

2017 ◽  
Vol 52 (4) ◽  
pp. 561-569 ◽  
Author(s):  
TC Sovernigo ◽  
PR Adona ◽  
PS Monzani ◽  
S Guemra ◽  
FDA Barros ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Embryo Production ◽  
Bovine Oocytes

Anethole improves blastocysts rates together with antioxidant capacity when added during bovine embryo culture rather than in the in vitro maturation medium

Zygote ◽  
2019 ◽  
Vol 27 (6) ◽  
pp. 382-385 ◽  
Author(s):  
J.C. Anjos ◽  
F.L.N. Aguiar ◽  
N.A.R. Sá ◽  
J.F. Souza ◽  
F.W.S. Cibin ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Cumulus Cells ◽  
Bovine Embryo ◽  
Oxygen Species ◽  
Embryo Production ◽  
Maturation Medium ◽  
Bovine Oocytes ◽  
Medium Supplementation

SummaryWe performed the exposure of bovine oocytes to anethole during in vitro maturation (0 or 300 µg/ml), during in vitro embryo production (0, 30, 300 or 2000 µg/ml), or during both periods to determine the rates of 2−4 cells embryos, blastocysts rates and cells numbers, as well as the production of reactive oxygen species (ROS). Bovine ovaries (n = 240) were collected from a local abattoir after slaughter and cumulus–oocyte complexes (COCs) with homogeneous and non-dark cytoplasm, surrounded by two or more compact layers of cumulus cells, and an intact zona pellucida were selected for in vitro maturatuion (IVM). Mature oocytes were then submitted to in vitro fertilization (IVF) and in vitro embryo production (IVP) in culture medium supplemented or not with different concentrations of anethole, as described above. Although IVM medium supplementation with 300 µg/ml anethole improved the rates of bovine blastocysts formation, we demonstrated that IVP medium supplementation with 30 µg/ml anethole, regardless of IVM medium enrichment, considerably enhanced blastocysts rates. Furthermore, ROS levels were decreased only when anethole was added to the IVP medium without previous IVM medium supplementation.

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