Gel-Loading Tip As Container for Vitrification of In Vitro-Produced Bovine Embryos.

Tominaga, Keiichiro; Hamada, Yukako

doi:10.1262/jrd.47.267

Cited by 35 publications

(33 citation statements)

References 21 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%

“…Newly developed cryodevices for accelerating the cooling rate include electron microscope grids (Martino et al, 1996), the open-pulled straw (OPS; Vajta et al, 1998), the Cryoloop (Lane et al, 1999) and the Cryotop (Kuwayama & Kato, 2000). Minor modifications of the container in the OPS system have also been reported (Kong et al, 2000;Hochi et al, 2001;Tominaga & Hamada, 2001). Hochi et al (2004) …”

Section: Introductionmentioning

confidence: 89%

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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%

Section: Introductionmentioning

confidence: 89%

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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“…However, it was difficult to control precisely the v ol um e of v i t r if i c at i on s olu t i on , p os s ib l y explaining the poor reproducibility of these ultrarapid vitrification procedures. It was examined whether commercially available gel-loading t ips (G L-Tips ) are available as containers for vitrification of IVP bovine embryos at various developmental stages [27], because the tip connected with micro-pipette allows the precise control of handling medium. Bovine embryos at D ay-1, -2, -3, -4, -5, and -7 i n 0.6-0.…”

Section: Ultra-rapid Vitrification Using Gl-tip As Containermentioning

confidence: 99%

“…Day 6.5-8 embryos were biopsied and then vitrified either by a GL-Tip method [27] or by a VSED-straw method [46] with minor modification. Pregnancy rate f o l l o w i n g t r a n s f e r o f s e x -i d e n t i f i e d a n d cryopreserved embryos in GL-Tip vitrification method (46.9%, 61/130) was higher than that in modified VSED-straw method (37.2%, 119/320), and was similar to that of fresh embryos (51.4%, 73/142).…”

Section: Cryopreservation Of Sex-identified Embryosmentioning

confidence: 99%

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Tominaga

2004

Journal of Reproduction and Development

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Abstract. My research awarded includes contributions to cryopreservation and sexing of bovine embryos produced in vitro and in vivo, as follows; (1) In vivo-derived morulae and blastocysts were cryopreserved in the presence of 10% glycerol, and the embryos were transferred into recipients after two-step dilution of glycerol in straw, with a practically acceptable pregnancy rate. (2) The survival rate of 16-cell stage embryos frozen in the medium with ethylene glycol was higher than that with DMSO or 1,2-propanediol. Addition of linoleic acid-albumin to culture medium enhanced the survival rate of post-thaw bovine 16-cell stage in vitro-produced (IVP) embryos. (3) Polarization of cytoplasmic lipid droplets by centrifugation of 2-cell stage embryos was found effective to increase freezing tolerance in 16-cell stage embryos developed from the centrifuged embryos, because blastomeres of 16-cell stage embryos were mostly lipid-free. (4) The usefulness of gel-loading tip (GLTip) as a container for ultra-rapid vitrification was demonstrated in IVP embryos from 2-cell to blastocyst stages, with a higher in vitro survival than the conventional two-step freezing. (5) PCR analysis for sexing of in vivo-derived Day-7 embryos indicated that male embryos developed faster and graded higher than female embryos. But such correlation between genetic sex and embryonic development was not found in IVP embryos obtained from individual cows. (6) Addition of 0.1-1.0% deproteinized hemodialysate product from calf blood to culture medium increased the producing efficiency of demi-embryos with good quality. Female embryos rather than male embryos required a longer time to repair after bisection. (7) In vivo-derived bovine embryos after biopsy for sexing by PCR analysis and subsequent vitrification using GL-Tips are available to practical use in the field. (8) Introduction of primer extension preamplification-PCR and purification of DNA product before standard sexing PCR of biopsy samples from Day 3-4 in vitro-derived embryos allowed accurate sex determination, and Day-7 blastocysts developed from Day 3-4 embryos were cryopreserved by GLTip vitrification without a loss of their viability. Thus the field application of bovine embryo transfer is in part supported by improvements of technologies in embryo cryopreservation and sex predetermination.

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“…Recently, ultra-rapid cooling methods have been developed using embryo containers, such as an electron microscope grid [5], an open pulled straw [3], a micro-glass pipette [6] and a gel loading tip [7], and by direct dropping of embryos into liquid nitrogen (LN 2 ) [8]. The viability of vitrifiedwarmed embryos has been greatly improved, however, these methods seem to be unsuitable for direct transfer because of contact of the solution containing embryos with the thawing solution (an open system).…”

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confidence: 99%

Birth of Lambs after Direct Transfer of Vitrified Ovine Embryos.

Okada

Wachi

Iida

et al. 2002

Journal of Reproduction and Development

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Abstract. The aim of this study was to investigate in vivo viability of vitrified ovine embryos after direct transfer using a laparoscope. In vivo-derived ovine morulae and blastocysts were equilibrated in 25% glycerol and 25% ethylene glycol solution, loaded into a heat-pulled straw and vitrified. Thirty-eight embryos were transferred into 19 recipients (2 embryos per recipient) and pregnancy was examined at 54-59 days after transfer. The pregnancy, implantation, lambing, and survival rates were 15.8% (3/19), 10.5% (4/38), 15.8% (3/19) and 7.9% (3/38), respectively. The result indicates that viable lambs can be produced by direct transfer of vitrified ovine embryos.

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Gel-Loading Tip As Container for Vitrification of In Vitro-Produced Bovine Embryos.

Cited by 35 publications

References 21 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

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Birth of Lambs after Direct Transfer of Vitrified Ovine Embryos.

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