Cryopreservation of in vitro produced bovine embryos: effects of lipid segregation and post-thaw laser assisted hatching

Pryor, J. H.; Looney, C.R.; Romo, S.; Kraemer, D.C.; Long, Charles R.

doi:10.1016/j.theriogenology.2010.07.006

Cited by 16 publications

(16 citation statements)

References 43 publications

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“…1). Although this method allows a good estimate of living and dead cells in the embryo, since dead cells are permeable to PI and living cells are stained by the Hoechst [8], an underestimation of dead cells may occur if they are surrounded by living cells [8].…”

Section: Resultsmentioning

confidence: 99%

“…An embryo's capacity to survive cryopreservation depends on several factors, including species, stage of development, origin (in vivo or in vitro), cryopreservation methodology, cryoprotectant [5], and intracytoplasmic lipid content [6]. Post-thaw viability can be estimated with the blastocoel reexpansion rate, embryonic cleavage rate, in vitro hatching [7,8], as well as with fluorescent probes (e.g., Hoechst 33 342 and propidium iodide), which stain living and dead cells, respectively, and are useful to estimate the percentage of viable blastomeres [8,9].…”

Section: Introductionmentioning

confidence: 99%

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Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

et al. 2012

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The objectives were to evaluate the reexpansion blastocoele rate, post-thaw viability, and in vitro development of canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol (GLY) or 1.5 m ethylene glycol (EG). Fifty-one in vivo-produced canine blastocysts were randomly allocated in two groups: GLY (n = 26) and EG (n = 25). After thawing, embryos from M0 were immediately stained with the fluorescent probes propidium iodide and Hoechst 33 342 to evaluate cellular viability. Frozen-thawed embryos from M3 and M6 were cultured in SOFaa medium + 10% FCS at 38.5°C under an atmosphere of 5% CO(2) with maximum humidity, for 3 and 6 days, respectively, and similarly stained. The blastocoele reexpansion rate (24 h after in vitro culture) did not differ between GLY (76.5%) and EG (68.8%). Post-thaw viable cells rate were not significantly different between GLY and EG (66.5 ± 4.8 and 57.3 ± 4.8, respectively, mean ± SEM), or among M0 (62.3 ± 5.7%), M3 (56.9 ± 6.0%), and M6 (66.5 ± 6.0%). In conclusion, canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol or 1.5 m ethylene glycol, had satisfactory blastocoele reexpansion rates, similar post-thawing viability, and remained viable for up to 6 days of in vitro culture.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

et al. 2012

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“…Many studies have tried to evaluate the effect of approaches such as increasing the cooling and re-warming speed with super-cooled nitrogen (WERLICH et al, 2006), use of cytoskeletal stabilizers (PRYOR et al, 2011), and use of different cryoprotectant combinations and reduced volumes of cryopreservation solution (TANIGUCHI et al, 2007;RIOS et al, 2010). Unfortunately, so far, none of the proposed strategies consistently improved the viability of bovine cryopreserved IVP embryos to the levels observed for in vivo-generated embryos.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro-produced (IVP) embryos are more sensitive to cryopreservation than their in vivoderived counterparts (LEIBO; LOSKUTOFF, 1993;PRYOR et al, 2011). IVP embryos differ from the in vivo-produced embryos in many aspects, including lower buoyant density (POLLARD; LEIBO, 1994), incomplete compaction (WOLFE; BRYANT, 1999), darker cytoplasm (DIEZ et al, 2001), and increased lipid levels (CROSIER et al, 2000;FAIR et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Effect of the cryopreservation method used, the embryonic stage and the use of conjugated linoleic acid isomers on the cryotolerance of in vitro-produced bovine embryos

et al. 2015

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Effect of the cryopreservation method used, the embryonic stage and the use of conjugated linoleic acid isomers on the cryotolerance of in vitro-produced bovine embryos Efeito do método de criopreservação utilizado, estágio de desenvolvimento embrionário e uso de isômeros do ácido linoleico conjugado na criotolerância de embriões bovinos produzidos in vitro AbstractConjugated linoleic acid (CLA) might be able to improve the cryotolerance of in vitro-produced (IVP) embryos. The effect of two CLA isomers on the cryotolerance of bovine IVP embryos, as well as that of the stage of embryonic development and the method used for cryopreservation was evaluated by three experiments. In Experiment 1, oocytes (n = 3,917) were fertilized in vitro and cultured with 0, 50, 100, or 200 µM trans-10, cis-12 (t10, c12 CLA). In Experiment 2, fertilized oocytes (n = 2,131) were cultured with 100 µM t10, c12 or cis-9, trans-11 (c9, t11 CLA), or a combination of both isomers. The embryos were vitrified at the blastocyst (BL) or the expanded blastocyst (EB) stage. In Experiment 3, oocytes (n = 1,720) were fertilized and cultured with or without 100 µM t10, c12 CLA, and the blastocysts were vitrified or frozen. Blastocyst development rate as well as the rates of re-expansion and hatching after thawing was recorded. Moreover, the mean cell number and mRNA expression of acetyl-CoA carboxylase (ACC1) and stearoyl-CoA desaturase (SCD1) as well as fatty acid synthase (FASN) multienzyme complex were determined. In Experiment 1, the highest concentration of t10, c12 CLA that did not reduce blastocyst development rate was 100 µM. In Experiment 2, the rates of re-expansion and hatching among the EBs obtained through IVP after supplementation with t10, c12 CLA (73.1% and 57.7%), with c9, t11 CLA (80.0% and 68.6%), with the combination (78.3% and 52.2%), and with the control group (85.4% and 58.3%) were similar. At the BL stage, the rates of re-expansion and hatching were lower than those at the EB stage, and CLA combination allowed a hatching rate (8.0%) lower than that observed in the control group (40.0%). In Experiment 3, the hatching rates for vitrified EBs (vitrified control; 67.4%) and vitrified CLA EBs (65.8%) were higher than those obtained for frozen EBs, exposed (13.3%) or not exposed (28.6%) to CLA. In addition, in Experiment 3, the hatching rate was higher at the EB stage in vitrified groups, while the rates of BL and EB were similar in frozen groups, thus proving that vitrification was more efficient than freezing for IVP bovine embryos. In Experiment 3, CLA isomer t10, C12 did not influence the embryonic cell number or mRNA expression of ACC1 and SCD1 enzymes, but decreased the mRNA expression of FASN. In conclusion, 100 µM CLA did not affect subsequent embryonic development. However, neither CLA isomer improved the cryotolerance of IVP bovine embryos. c12 CLA). No Experimento 2, oócitos fecundados (n = 2.131) foram cultivados com 100 µM de t10, c12 ou cis-9, trans-11 (c9, t11 CLA), ou ainda com uma associação de ambos. Os embri...

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“…From the early days of IVF, it was obvious that in-vitromatured oocytes and in-vitro-produced embryos looked and behaved differently. For example, several studies show that in-vitro-produced embryos are less competent to implant and more sensitive to cryopreservation compared to in-vivo-produced embryos (Pryor et al 2011). With the development of genomic tools, the capacity to analyse embryos from the one-cell to blastocyst stage provided a new angle to compare in vivo and in vitro aspects.…”

Section: The Genomic Effect (2010-2020)mentioning

confidence: 99%

40 years of bovine IVF in the new genomic selection context

Sirard¹

2018

Reproduction

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The development of a complex technology such as fertilization (IVF) requires years of experimentation, sometimes comparing several species to learn how to create the right environment for oocytes, spermatozoa and early embryos. At the same time, individual species characteristics such as gamete physiology and gamete interaction are recently evolved traits and must be analysed within the context of each species. In the last 40 years since the birth of Louise Brown, IVF techniques progressed and are now used in multiple domestic and non-domestic animal species around the world. This does not mean that the technology is completely matured or satisfactory; a number of problems remain to be solved and several procedures still need to be optimized. The development of IVF in cattle is particularly interesting since agriculture practices permitted the commercial development of the procedure and it is now used at a scale comparable to human IVF (millions of newborns). The genomic selection of young animals or even embryos combined with sexing and freezing technologies is driving a new era of IVF in the dairy sector. The time has come for a retrospective analysis of the success and pitfalls of the last 40 years of bovine IVF and for the description of the challenges to overcome in the years to come.

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Cryopreservation of in vitro produced bovine embryos: effects of lipid segregation and post-thaw laser assisted hatching

Cited by 16 publications

References 43 publications

Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

Effect of the cryopreservation method used, the embryonic stage and the use of conjugated linoleic acid isomers on the cryotolerance of in vitro-produced bovine embryos

40 years of bovine IVF in the new genomic selection context

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