In vitro development following one-step dilution of OPS-vitrified porcine blastocysts

Cuello, C.; Gil, M.A.; Parrilla, Inmaculada; Tornel, Jose; Vázquez, Juan M.; Roca, Jordi; Berthelot, F.; Martinat-Botté, F.; Martı́nez, Emilio A.

doi:10.1016/j.theriogenology.2003.12.018

Cited by 63 publications

(30 citation statements)

References 20 publications

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“…This good survival rate was not due solely to the cryoprotectant combination, but also to the multistep dilution in the warming procedure. This multistep dilution has an important role in the prevention of sudden osmolarity changes that could happen during the one step dilution method [43].…”

Section: Discussionmentioning

confidence: 99%

In vitro maturation, apoptotic gene expression and incidence of numerical chromosomal abnormalities following cryotop vitrification of sheep cumulus-oocyte complexes

Ebrahimi

Valojerdi

Eftekhari‐Yazdi

et al. 2010

J Assist Reprod Genet

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Purpose The purpose of this study was to evaluate the effects of cryotop vitrification of sheep cumulus-oocyte complexes (COCs) on oocyte maturation, apoptotic gene expression and incidence of chromosomal abnormalities. Methods Freshly isolated (control group) and vitrifiedwarmed COCs (cryotop group) were matured in vitro. The expression of pro-and anti-apoptotic genes was investigated by real-time PCR. The incidence of numerical chromosomal abnormalities was evaluated by cytogenetic analysis. Results The mean percentage of oocytes in the cryotop and control groups that reached metaphase II was 49.25±3.01% and 51.94±2.7% respectively. The expression rates of proand anti-apoptotic genes were similar in both groups, whereas the incidence of numerical chromosomal abnormalities was higher in the cryotop group compared to the control group (42.5% vs. 20%, p<0.05). Conclusion Although cryotop vitrification of COCs did not affect the incidence of oocyte maturation or apoptotic gene expression, significant deficiencies in the maintenance of oocyte chromosomal organization were seen.

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Section: Discussionmentioning

confidence: 99%

In vitro maturation, apoptotic gene expression and incidence of numerical chromosomal abnormalities following cryotop vitrification of sheep cumulus-oocyte complexes

Ebrahimi

Valojerdi

Eftekhari‐Yazdi

et al. 2010

J Assist Reprod Genet

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“…Since vitrification uses CPAs at high concentrations to prevent intracellular ice crystal formation, their toxicity causes damage to embryos when the vitrification procedure takes a long time. To avoid this problem, several vitrification methods, such as the openpulled straw [5], microdroplet [6] and metal mesh vitrification [7] methods, have been reported to achieve high viability by making porcine embryos very rapidly pass a critical temperature range at which embryos are injured. The viability of embryos is increased by these methods, as they have the advantages of minimizing the required amounts of CPAs by using special devises and lowering the concentration of CPAs by application of rapid cooling.…”

Section: Discussionmentioning

confidence: 99%

“…Due to the extremely high sensitivity of porcine embryos to low temperature, it has been difficult with slow freezing methods to achieve conception rates and litter sizes equivalent to those achieved with artificial insemination [1,2]. However, it has recently become possible to produce piglets from cryopreserved embryos by using ultra-rapid vitrification methods, such as the minimum volume cooling (MVC) [3,4], open pulled straw [5], microdroplet [6] and metal mesh vitrification [7] methods.…”

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confidence: 99%

Viability of Porcine Embryos after Vitrification Using Water-soluble Pullulan Films

Sakagami¹,

Yamamoto²,

Akiyama³

et al. 2010

Journal of Reproduction and Development

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Abstract. The efficiency of a porcine embryo vitrification method that uses water-soluble films of pullulan, a naturallyoccurring polysaccharide polymer, was compared with two other types of vitrification methods using different devices and solutions for vitrification and warming. Blastocysts collected in vivo and vitrified by the conventional straw (ST), Cryotop ® (MVC) or pullulan film vitrification (PFV) methods were stored in liquid nitrogen for a certain period of time, after which the cryoprotective agents were removed by stepwise dilution. Fresh embryos were used as controls for the non-vitrification group. The vitrified-warmed embryos were incubated in TCM199 with 0.1 mM β-mercaptoethanol and 20% fetal bovine serum for 24 h at 38.5 C in humidified air with 5% CO2 to evaluate their viability. The survival rate of embryos in the ST group (48.3%) was significantly lower than that of those in the MVC (70.7%), PFV (79.0%) and nonvitrification (94.4%) groups. The oxygen consumption rate after vitrification was significantly lower than that before vitrification in the ST group, but was not significantly different in the MVC and PFV groups. Both the oxygen consumption rates of embryos after warming and the live cell numbers in the ST group were lower than those in the MVC group, while they did not differ significantly between the PFV and MVC groups. There was a correlation between the oxygen consumption rate and the number of live cells in vitrified embryos after warming. Our results demonstrated that in vivo-derived porcine embryos could be vitrified using pullulan films. Key words: Oxygen consumption rate, Porcine embryo, Pullulan film, Vitrification (J. Reprod. Dev. 56: [279][280][281][282][283][284] 2010) reservation of porcine embryos is important for increasing the effective use of high-quality genetic resources, preventing disease transmission via animals and allowing low-cost transportation of pigs. Due to the extremely high sensitivity of porcine embryos to low temperature, it has been difficult with slow freezing methods to achieve conception rates and litter sizes equivalent to those achieved with artificial insemination [1,2]. However, it has recently become possible to produce piglets from cryopreserved embryos by using ultra-rapid vitrification methods, such as the minimum volume cooling (MVC) [3,4], open pulled straw [5], microdroplet [6] and metal mesh vitrification [7] methods.Pullulan, which is a neutral polysaccharide polymer also known as α-1,4-; α-1,6-glucan, is made from starch and consists of maltotriose units linked in an orderly manner. It has been reported that murine morulae placed on a pullulan film could be vitrified [8]. In general, stepwise dilution is required after warming of vitrified embryos, since vitrification requires a high concentration of cryoprotective agents (CPAs), which makes it difficult, due to their toxicity, to warm the embryos in a straw for direct transfer to recipients. As the pullulan film is soluble in warm water, the vitrification solution can be diluted ...

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“…Vitrification of untreated morulae and blastocysts has resulted in high survival rates after warming (Berthelot et al 2003), especially when re-warming after SOPS is done in one stage (direct warming, a very practical solution for ET, Cuello et al 2004b), yielding live litters . For blastocysts (See Figure 5), use of the SOPS waived the need for centrifugation (dislocation of lipids) or microtubule stabilization, thus making the method a very practical one and indicating the procedure is now reaching maturity for commercial application (Cameron et al 2004, Beebe et al 2005, Martinat-Botté et al 2006, Cuello et al 2008, Sanchez-Osorio et al 2009.…”

Section: Cryopreservation Of Oocytes and Embryosmentioning

confidence: 99%

Cryopreservation of Porcine Gametes, Embryos and Genital Tissues: State of the Art

Rodriguez‐Martinez¹

2012

Current Frontiers in Cryobiology

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In vitro development following one-step dilution of OPS-vitrified porcine blastocysts

Cited by 63 publications

References 20 publications

In vitro maturation, apoptotic gene expression and incidence of numerical chromosomal abnormalities following cryotop vitrification of sheep cumulus-oocyte complexes

In vitro maturation, apoptotic gene expression and incidence of numerical chromosomal abnormalities following cryotop vitrification of sheep cumulus-oocyte complexes

Viability of Porcine Embryos after Vitrification Using Water-soluble Pullulan Films

Cryopreservation of Porcine Gametes, Embryos and Genital Tissues: State of the Art

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