Cellular and molecular characterization of the impact of laboratory setup on bovine in vitro embryo production

Plourde, Dany; Vigneault, Christian; Laflamme, Isabelle; Blondin, Patrick; Robert, Claude

doi:10.1016/j.theriogenology.2011.12.021

Cited by 15 publications

(13 citation statements)

References 57 publications

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“…In the present study, control and ROS groups had more apoptotic cells in comparison with the groups that were treated with BL-I and ROS + BL-I. These results differ from other studies, where the total of apoptotic cells was similar between the control group and groups treated with inhibitors (Lonergan et al 2000;Mermillod et al 2000;Adona et al 2008;Plourde et al 2012).…”

Section: Discussioncontrasting

confidence: 99%

“…In the present study, differences in the number of embryo viable cells were observed when comparing different groups. Plourde et al (2012) comparing two IVP laboratories (A and B) observed that embryos from laboratory A had an average of 111 AE 23 embryonic cells and embryos from laboratory B had an average of 115 AE 25. In the present study, embryos from control and ROS groups showed similar results to those obtained by these authors (108.2 and 109.7 embryonic cells, respectively).…”

Section: Discussionmentioning

confidence: 99%

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Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification

Maziero

Guaitolini

Paschoal

et al. 2016

Reprod Domestic Animals

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This study aimed to produce in vitro bovine embryos by the addition of two drugs, which is responsible for oocyte meiosis inhibition: roscovitine (ROS) and butyrolactone I (BL-I). Oocytes were recovered from slaughtered cows and matured in a commercial medium and maintained in a 5% CO2 atmosphere. Oocytes were maintained for 6 h in an in vitro maturation (IVM) medium containing ROS (12.5 μm), BL-I (50 μm) and association of drugs (ROS 6.25 μm and BL-I 25 μm). Oocytes were cultured for 18 h in an agent-free medium for the resumption of meiosis. After 24 h of maturation, oocytes were inseminated in the commercial in vitro fertilization (IVF) medium. Presumptive zygotes were cultured in SOFaa medium in a 5% CO2 atmosphere. On day 3, rate of cleavage was evaluated and on days 6 and 7, rate of blastocyst formation. BL-I and its association with the ROS increased the rates of cleavage and blastocyst formation (p < 0.05). The ROS alone was inefficient, impairing embryonic development, with low rates of blastocyst formation when compared to the control group and other treatments (p < 0.05). The embryos from BL-I and ROS+BL-I groups presented higher number of cells and lower rates of cellular apoptosis compared to other groups, either for the fresh or for post-thawing embryos. Embryos from ROS+BL-I group showed to be more resistant to the vitrification process, presenting a higher rate of embryonic re-expansion (p < 0.05). In conclusion, block of meiosis using BL-I or its association with ROS increased the rate of blastocyst formation, and the association of ROS+BL-I resulted in a better resistance to the embryo cryopreservation process.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification

Maziero

Guaitolini

Paschoal

et al. 2016

Reprod Domestic Animals

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“…The ART procedures that can be applied to enhance reproduction in this industry are fertilization of oocytes either in vitro through collection of oocytes or in vivo through artificial insemination, embryo recovery through uterine flushing, in vitro embryo production and transfer of embryos to surrogates (Wu, 2012). In vitro embryo production through its processes such as oocytes maturation, fertilization and subsequent embryo production allows more frequent gametes collection and therefore further increase the number of transferable embryos (Plourde, 2012). Additionally, it has been mention that in vitro embryo production technique is estimated to yield three times more embryos than other ARTs through the constant collection of oocytes (Plourde, 2012).…”

Section: Overview Of In Vitro Embryo Production In the Beef Cattle Inmentioning

confidence: 99%

“…In vitro embryo production through its processes such as oocytes maturation, fertilization and subsequent embryo production allows more frequent gametes collection and therefore further increase the number of transferable embryos (Plourde, 2012). Additionally, it has been mention that in vitro embryo production technique is estimated to yield three times more embryos than other ARTs through the constant collection of oocytes (Plourde, 2012). Retrieval of immature oocytes from cattle for initialising IVP can either be achieved from slaughter house ovaries or from a live donor cow.…”

Section: Overview Of In Vitro Embryo Production In the Beef Cattle Inmentioning

confidence: 99%

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145 the Role of Cell Apoptosis on in Vitro-Produced Beef Cattle Embryos

Nkadimeng

2017

Reprod. Fertil. Dev.

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Successful in vitro development of embryos is dependent upon maintenance of cellular function in the embryonic microenvironment. Exposure of pre-implantation embryos to a variety of cellular stresses can induce apoptosis in all or a fraction of blastomeres. Among the conditions that can induce apoptosis in cattle embryo production in vitro are heat shock, media composition, oxygen levels, and tumour necrosis factor (TNF)-α. The aim of the study was therefore to evaluate the in vitro culture environment of embryos in terms of cell apoptosis through caspase and DNA fragmentation evaluation. A total of 1200 immature oocytes were collected at slaughter from indigenous South African cow ovaries. The cumulus-oocyte complexes were randomly allocated into 2 maturation temperatures, 39°C and 41°C (200/temperature /replicated 6 times), and cultured in M199 + FSH-LH-oestradiol medium under oil at 100% humidity and 5% CO2 for 24 h. Postmaturation, oocytes were subjected to normal subsequent embryo conditions (Rynkowska et al. 2011 Biotechnol. Comput. Biol. Bionanotechnol. 92, 45–53). All matured oocytes were fertilized for 6 h with frozen-thawed Nguni bull semen from 10 bulls replicated 5 times at a concentration of 265 × 106 sperm cells/mL in Bracket and Oliphant medium under oil. The presumptive zygotes from each treatment were cultured into SOF-BSA medium under oil and incubated at 39°C for assessment of cleavage rate post IVF. Produced embryos were divided into 3 groups: 2–4 cell, ≥8 cell embryos, and blastocyst. Two- to four-cell embryos were removed at Day 2, ≥8 cell at Day 5 (mainly 8-cell, morula, and cavitated blastocyst), and expanded blastocyst at Day 7 of embryo culture to determine caspase activity and DNA fragmentation for evidence of apoptosis. Caspase was performed using colourimetric assay on a 96-well microplate reader and monitored at 450 nm reference filter. The DNA fragmentation was examined using the TUNEL assay method and imaging was done using the Alexa Fluor® 488 emission at 519 nm. Data were analysed using SAS 9.2 software (SAS Institute Inc., Cary, NC, USA), and Shapiro-Wilks test was used on the standardized residuals to test for deviations from normality, means of significant effects were compared using Student’s t-l.s.d. at the 95% confidence interval. There was no significant difference on the cleavage rates [39°C (72.0 ± 22.70) and 41°C (67.2 ± 18.9)] and blastocysts [39°C (11.4 ± 2.6) and 41°C (11.2 ± 6.3)] from both maturation temperatures. Caspase activity showed no significant difference on 2–4 cell [39°C (0.015 ± 0.001) and 41°C (0.016 ± 0.002)] and ≥8 cell embryos at both temperatures [39°C (0.022 ± 0.007a) and 41°C (0.032 ± 0.013)]. However, blastocyst differed significantly at both temperatures [39°C (0.037 ± 0.012) and 41°C (0.053 ± 0.005)]. A higher (P < 0.05) DNA fragmentation was observed at 2–4 cell (6.5 ± 2.9), ≥8 cell (13.3 ± 3.1), and blastocyst (48.0 ± 8.2) for embryos produced from 41°C matured oocytes compared to the 39°C maturation group [2–4 cell (2.2 ± 1.2), ≥8 cell (4.5 ± 1.9), and blastocyst (9.7 ± 6.7)]. It is therefore concluded that embryos produced from 41°C matured oocytes can have the same developmental capacity as the 39°C maturation group. Moreover, both temperatures can show signs of apoptosis, however, with more apoptosis evidence in the 41°C than the 39°C maturation group.

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Maternal-embryo interaction in the bovine oviduct: Evidence from in vivo and in vitro studies

et al. 2016

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Cellular and molecular characterization of the impact of laboratory setup on bovine in vitro embryo production

Cited by 15 publications

References 57 publications

Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification

Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification

145 the Role of Cell Apoptosis on in Vitro-Produced Beef Cattle Embryos

Maternal-embryo interaction in the bovine oviduct: Evidence from in vivo and in vitro studies

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