Cryopreservation of Porcine Embryos Derived from In Vitro-Matured Oocytes1

Esaki, Ritsuko; Ueda, Hideto; Kurome, Mayuko; Hirakawa, Kazumasa; Tomii, Ryo; Yoshioka, Hiroki; Ushijima, Hitoshi; Kuwayama, Masashige; Nagashima, Hiroshi

doi:10.1095/biolreprod.103.026542

Cited by 90 publications

(64 citation statements)

References 30 publications

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“…However, in the present study, the survivability of GV-stage minke whale oocytes was improved by applying minimum volume cooling methods, such as the Cryotop and OPS methods. The Cryotop and OPS methods have been used for oocytes and/or embryos in cattle (Vajta et al, 1998;Tominaga & Hamada, 2001), pigs (Esaki et al, 2004), rabbits (Hochi et al, 2004) and humans (Kuwayama & Kato, 2000). The proportion of post-warm oocytes with normal morphology was significantly higher when the Cryotop rather than the OPS was used as the cryodevice, and a maximum proportion of post-warm oocytes extruding the first PB after IVM of 29.1% was obtained.…”

Section: Discussionmentioning

confidence: 99%

Effects of cryodevice type and donors' sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage

Iwayama

Hochi

Kato

et al. 2004

Zygote

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SummaryGerminal-vesicle-stage oocytes enclosed with compact cumulus cell layers (COCs) were recovered from adult or prepubertal minke whale ovaries, and were vitrified in a solution containing 15% ethylene glycol, 15% DMSO and 0.5 M sucrose using either a Cryotop or an open-pulled straw (OPS) as the cryodevice. The post-warm COCs with normal morphology were cultured for 40 h in a 390 mosmol in vitro maturation medium, and oocytes extruding the first polar body were considered to be matured. The proportion of morphologically normal COCs after vitrification and warming was higher when the COCs were cryopreserved by Cryotop (adult origin, 88.4%; prepubertal origin, 80.8%) compared with the OPS (adult origin, 67.7%; prepubertal origin, 64.2%). The oocyte maturation rate was higher in the adult/Cryotop group (29.1%) compared with those of the prepubertal/Cryotop group (14.4%), the adult/OPS group (14.3%) and the prepubertal/OPS group (10.6%). These results indicate that the Cryotop is a better device than the OPS for vitrification of immature oocytes from adult minke whales.

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Section: Discussionmentioning

confidence: 99%

Effects of cryodevice type and donors' sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage

Iwayama

Hochi

Kato

et al. 2004

Zygote

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“…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].…”

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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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“…Dinnyés et al (2000) reported that blastocyst development of vitrified bovine oocytes by SSV was 20%. Esaki et al (2004) cryopreserved porcine embryos by the MVC technique; the survival rate was 41%. Rojas et al (2004) obtained 14% cleavage rate after OPS vitrification of porcine oocytes.…”

Section: Discussionmentioning

confidence: 99%

Vitrification of in vitro matured oocytes of Mangalica and Large White pigs

Varga

Gajdócsi

Makkosné

et al. 2008

Acta Veterinaria Hungarica

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The breeding of Mangalica, a native pig breed in Hungary, had been started in 1833, but this pig breed almost became extinct in Hungary in the past decades. In 1991, the number of sows was only 200. Although in these days the existing Mangalica population consists of more than 6000 animals representing different colour variations, the preservation of this traditional pig breed is still very important. Vitrification is a potential tool for the preservation of gametes and embryos of these animals. The aim of this study was to investigate the effects of vitrification on the developmental competence of Mangalica (M) and Large White (LW) oocytes following fertilisation. The oocytes were vitrified by the Open Pulled Straw (OPS) method using different concentrations of ethylene glycol and dimethyl sulphoxide as cryoprotectants. After rehydration the oocytes underwent in vitro fertilisation; the resultant zygotes were then cultured in vitro for four days to assess embryonic development. In the first experiment, in vitro maturation of M and LW oocytes was compared. No significant difference was observed in the nuclear maturation rate of LW and M oocytes. In the second experiment, the sensitivity of oocytes to vitrification was examined by evaluating oocyte morphology after thawing. A higher percentage of LW oocytes showed normal morphology compared to M oocytes, indicating that Mangalica oocytes are more sensitive to cryoprotectants than Large White oocytes. After warming and in vitro fertilisation, more than 50% of the oocytes started embryonic development and by the end of the incubation period morula stage embryos had developed in both groups. The results show that the OPS vitrification technique is well suited to preserve Mangalica oocytes and from these oocytes morula embryos can be produced.

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Cryopreservation of Porcine Embryos Derived from In Vitro-Matured Oocytes1

Cited by 90 publications

References 30 publications

Effects of cryodevice type and donors' sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage

Effects of cryodevice type and donors' sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage

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

Vitrification of in vitro matured oocytes of Mangalica and Large White pigs

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