Effect of MEM vitamins and forskolin on embryo development and vitrification tolerance of in vitro-produced pig embryos

The aim of this study was to evaluate the effects of the two types of media, namely minimum essential medium (αMEM) and Ham'sF10, supplemented with foetal bovine serum (FBS) or bovine serum albumin (BSA) in vitrification/warming solution on the quality and viability of sheep ovarian follicles. Vitrification method was applied for cryopreservation of sheep ovarian cortex using Ham'sF10 and αMEM supplemented with either BSA or FBS. There were five groups: Fresh, Ham'sF10+ BSA, Ham'sF10+ FBS, αMEM + BSA and αMEM + FBS. Samples were cultured for two weeks after warming. Viability and morphology of follicles and DNA fragmentation in follicles and in tissue stroma cells were analysed before vitrification/warming and following one and two weeks of culture. The Ham'sF10+ FBS and Ham'sF10+ BSA groups showed a significant decrease in follicular viability after one week of culture (p < .05 vs. Fresh). Following two weeks of culture, all groups revealed a considerable fall in the number of viable follicles (p < .05 vs. Fresh). There was an increase in DNA fragmentation of connective tissue cells but not in the follicles (p < .05). Our results showed the better application of αMEM supplemented with BSA as a vitrification solution in improvement of cryopreservation effects and maintenance of follicular survival.

Section: Discussionmentioning

confidence: 91%

Comparison of the effects of Ham'sF10 and αMEM in combination with FBS or BSA in vitrification/warming solution on quality and viability of sheep ovarian follicles

Mohammadzadeh

Safdarian

Amidi

et al. 2017

“…Age and parity influence reproductive performance after artificial insemination (AI), with gilts and primiparous sows being less efficient than multiparous sows (Clark, Schinckel, Singleton, Einstein, & Teclaw, ). Multiparous sows are ideal as recipients because the insertion of the NsDU‐ET catheter is easier, their conduct during the transfers is better (Martinez, Cuello et al., ), and their oestruses are naturally and effectively synchronized by managing weaning. All of the abovementioned observations led us to evaluate the influence of recipient parity (parity 1 to 5) on NsDU‐ET outcomes (Martinez, Nohalez et al., ).…”

Section: Factors Affecting the Et Technologymentioning

confidence: 99%

“…We also worked on intracytoplasmic lipids at several levels. In vivo‐derived zygotes treated with forskolin, a lipolytic agent that increased the survival of in vitro‐derived blastocysts (Cuello et al., ), prior to vitrification enhanced their vitrification ability, resulting in a post‐warming blastocyst formation rate of 75% (Gomis et al., 2013b). In addition, lipid polarization pre‐treatments before vitrification also increased the post‐warming capacity of in vivo‐derived zygotes to reach the blastocyst stage (Gomis et al., 2013a).…”

Section: Researches On Embryo Vitrificationmentioning

confidence: 99%

Achievements and future perspectives of embryo transfer technology in pigs

Martı́nez

Martínez

Cambra

et al. 2019

Commercial embryo transfer (ET) has unprecedented productive and economic implications for the pig sector. However, pig ET has been considered utopian for decades mainly because of the requirements of surgical techniques for embryo collection and embryo deposition into recipients, alongside challenges to preserve embryos. This situation has drastically changed in the last decade since the current technology allows non-surgical ET and short-and long-term embryo preservation. Here, we provide a brief review of the improvements in porcine ET achieved by our laboratory in the past 20 years. This review includes several aspects of non-surgical ET technology and different issues affecting ET programmes and embryo preservation systems.The future perspectives of ET technology are also considered. We will refer only to embryos produced in vivo since they are the only type of embryos with possible short-term use in pig production. K E Y W O R D Sembryo storage, embryo transfer, porcine, vitrification | 5 MARTINEZ ET Al.

“…Moreover, few studies had vitrified embryos after meiotic inhibition, and only Ponderato et al (2001) report that having cryopreserved embryos after the use of ROS + BL-I (12.5 lM ROS + 6.25 lM BL-I) for 24 h obtained similar data between control and treated groups. Cuello et al (2013) when comparing fresh embryos to vitrified and heated embryos observed a similarity in the total of cells; however, Dhali et al (2011) found a higher number of cells in fresh embryos than in vitrified ones (157 and 130, respectively). These results differ from those obtained in the present study, where all groups of fresh embryos had a higher number of intact cells, when compared to the number of intact cells from the vitrified embryo groups (Table 5).…”

Section: Discussionmentioning

confidence: 95%

“…Cuello et al. () when comparing fresh embryos to vitrified and heated embryos observed a similarity in the total of cells; however, Dhali et al. () found a higher number of cells in fresh embryos than in vitrified ones (157 and 130, respectively).…”

Section: Discussionmentioning

confidence: 98%

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

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.