Field trial to compare pregnancy rates of bovine embryo cryopreservation methods: Vitrification and one-step dilution versus slow freezing and three-step dilution

Leeuw, A.M. van Wagtendonk-de; Daas, J.H.G. den; Rall, W.F.

doi:10.1016/s0093-691x(97)00340-3

Cited by 51 publications

(24 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This may be partially due to the high developmental competence of in vivo-produced embryos compared with IVP embryos. Previously, van Wagtendonk-de Leeuw et al [8] reported that in field trials, vitrification does not appear to be more efficient than conventional slow freezing for cryopreservation of in vivo-derived bovine embryos in terms of pregnancy. Thus, our vitrification method using Cryotop also may not achieve higher rates of pregnancy after ET of cryopreserved in vivo-derived bovine embryos than the conventional freezing method.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, it is difficult to perform vitrification in the field. Recently, however, some vitrification methods have been reported as suitable for in-straw dilution of embryos; these methods may simplify the ET procedure to such an extent that it may be used to transfer vitrified embryos on farms at the same level of complexity as carrying out an artificial insemination with frozen-thawed semen [8,11,12].To facilitate vitrification, several methods that require very small sample volumes have been devised. For example, decreasing the sample volume to <1 μl and using carrier systems of minimum capacity increase the heat conduction of the sample, which can then be cooled very quickly.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

“…Vitrification reduces the time commitment and equipment expense associated with cryopreservation compared with conventional slow freezing [7]. However, vitrification has not been widely adopted by ET practitioners for commercial use in cattle [7,8]. In general, vitrification requires gradual cryoprotectant dilution in a laboratory setting [9,10] because it uses a high concentration of cryoprotectants.…”

mentioning

confidence: 99%

See 3 more Smart Citations

In-straw Cryoprotectant Dilution for Bovine Embryos Vitrified Using Cryotop

et al. 2011

View full text Add to dashboard Cite

Abstract. The aim of this study was to develop an in-straw dilution method suitable for 1-step bovine embryo transfer of vitrified embryos using the Cryotop vitrification-straw dilution (CVSD) method. The development of embryos vitrified using the CVSD method was compared with those of embryos cryopreserved using in-straw vitrificationdilution (ISVD) and conventional slow freezing, outside dilution of straw (SFODS) methods. In Experiment 1, in vitroproduced (IVP) embryos cryopreserved using the CVSD method were diluted, warmed and exposed to the dilution solution at various times. When vitrified IVP embryos were exposed to the dilution solution for 30 min after warming, the rates of embryos developing to the hatched blastocyst stage after 72 h of culture (62.0-72.5%) were significantly lower (P<0.05) than those of embryos exposed to the solution for 5 and 10 min (82.4-94.3%), irrespective of supplementation with 0.3 M sucrose in the dilution solution. In Experiment 2, the rate of embryos developing to the hatching blastocyst stage after 48 h of culture in IVP embryos cryopreserved using the SFODS method (75.0%) was significantly (P<0.05) lower than those of embryos cryopreserved using the CVSD and ISVD methods (93.2 and 97.3%, respectively). In Experiment 3, when in vivo-produced embryos that had been cryopreserved using the CVSD, ISVD and SFODS methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception and delivery rates among groups. In Experiment 4, when IVP embryos derived from oocytes collected by ovum pick-up that had been cryopreserved using the CVSD and ISVD methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception rates among groups. Our results indicate that this simplified regimen of warming and diluting Cryotop-vitrified embryos may enable 1-step bovine embryo transfer without the requirement of a microscope or other laboratory equipment. Key words: Bovine, Conception rate, Cryotop, Embryo transfer, Vitrification (J. Reprod. Dev. 57: [437][438][439][440][441][442][443] 2011) ince the first successful cryopreservation of bovine embryos [1], cryopreservation of bovine embryos has been widely used commercially. A recent worldwide inventory revealed that more than 250,000 bovine in vivo-produced embryos have been used for embryo transfer (ET) after freezing and thawing [2]. However, the pregnancy rate of frozen-thawed embryos is slightly lower than that of fresh embryos [3]. And the pregnancy rate of frozen-thawed in vitro-produced (IVP) embryos is also significantly lower than that of fresh IVP embryos. Therefore, it is necessary to develop an embryo cryopreservation method to obtain higher conception rates. Recent reports have confirmed that vitrification of embryos, especially IVP embryos, is at least as efficient as conventional slow freezing [4][5][6][7]. Vitrification reduces the time commitment and equipment expense associated with cryopreservation compared with con...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

In-straw Cryoprotectant Dilution for Bovine Embryos Vitrified Using Cryotop

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Direct transfer of cryopreserved-thawed embryos reduces the time and expense involved in embryo transfer procedures and it offers handling advantages over conventional freezing procedures, which require step-wise removal of the cryoprotectant after thawing [ 10,11 ]. Under Embryos were frozen in 10 % glycerol in PBS using a conventional freezing method [8].…”

mentioning

confidence: 99%

Effect of post-thaw cell rehydration at 4 °C on survival of frozen and vitrified IVP-derived bovine embryos

Gutiérrez‐Adán¹,

Granados²,

Fuente³

1999

Reprod. Nutr. Dev.

View full text Add to dashboard Cite

-In vitro-produced bovine embryos (IVP) were either frozen in 10 % glycerol in a phosphate-buffered saline solution (PBS) using conventional slow freezing or vitrified in 25 % glycerol and 25 % ethylene glycol in PBS. The results of viability and hatching rates were compared between frozen and vitrified embryos after thawing and dilution using one of three different protocols: (A) a three-step dilution procedure, (B) a one-step dilution procedure or (C) a procedure in which embryos were kept in situ inside the straw at 4 °C for 10 min during a one-step dilution procedure. No

show abstract