Vitrification of human 8-cell embryos, a modified protocol for better pregnancy rates

Raju, Gottumukkala Achyuta Rama; Haranath, G. B.; Krishna, Kota Murali; Prakash, G. Jaya; Madan, K.

doi:10.1016/s1472-6483(10)61135-2

Cited by 109 publications

(22 citation statements)

References 23 publications

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“…In the study of Rama Raju et al (2005) [28], cleavage stage embryos were vitrified using opened devices or conventional slow-freezing protocol. Post-thaw survival rate was significantly higher for the vitrification group compared to the slow-freezing group ( p < 0.05).…”

Section: Discussionmentioning

confidence: 99%

“…Rama Raju et al (2005) showed a higher IR in vitrification group compared to slow-freezing group of patients [28]. Later, Balaban et al (2008) [29] confirmed a high clinical implantation rate after transfer of vitrified-warmed embryos.…”

Section: Discussionmentioning

confidence: 99%

“…It has to be noted that improved slow-freezing methods can produce comparable results [26, 27]. However, there are still only few randomized clinical trials addressing the optimal method of cryopreservation for human cleavage stage embryos [28–30]. …”

Section: Introductionmentioning

confidence: 99%

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A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos

Fasano

Fontenelle

Vannin

et al. 2013

J Assist Reprod Genet

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PurposeTo compare two different vitrification methods to slow freezing method for cryopreservation of human cleavage stage embryos. Design: Prospective randomised trial. Setting: University assisted reproduction centre. Patient(s): 568 patients (mean age 33.4 ± 5.2) from April 2009 to April 2011.Methods1798 supernumerary good-quality cleavage stage embryos in 645 IVF cycles intended to be cryopreserved were randomly allocated to three groups: slow freezing, vitrification with the Irvine® method, vitrification with the Vitrolife® method. Main Outcome Measure(s): Embryo survival and cleavage rates, implantation rate.ResultsA total of 1055 embryos were warmed, 836 (79.2 %) survived and 676 were finally transferred (64.1 %). Post-warming embryos survival rate was significantly higher after vitrification (Irvine: 89.4 %; Vitrolife: 87.6 %) than after slow freezing (63.8 %) (p < 0.001). No differences in survival rates were observed between the two vitrification methods, but a significant higher cleavage rate was observed using Irvine compared to Vitrolife method (p < 0.05). Implantation rate (IR) per embryo replaced and per embryo warmed were respectively 15.8 % (41/259) and 12.4 % (41/330) for Irvine, 17.0 % (40/235) and 12.1 % (40/330) for Vitrolife, 21.4 % (39/182) and 9.9 % (39/395) for slow-freezing (NS).ConclusionsBoth vitrification methods (Irvine and Vitrolife) are more efficient than slow freezing for cryopreservation of human cleavage stage embryos in terms of post-warming survival rate. No significant difference in the implantation rate was observed between the three cryopreservation methods.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos

Fasano

Fontenelle

Vannin

et al. 2013

J Assist Reprod Genet

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“…Currently, two basic techniques are used in human embryo cryopreservation: slow freezing and vitrification [4,5]. Both techniques can affect the developmental competence and function of embryos, as the embryos are exposed to various stressors during cryopreservation.…”

Section: Introductionmentioning

confidence: 99%

In vitrodevelopment and gene expression of frozen-thawed 8-cell stage mouse embryos following slow freezing or vitrification

Shin

Choi

Kim

et al. 2011

Clin Exp Reprod Med

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ObjectiveThis study was performed to compare the efficiency of slow freezing and vitrification based on survival, development to blastocysts, and cell numbers of blastocysts. Changes in embryonic gene expression in fresh and frozen-thawed embryos were also examined.MethodsEight-cell stage embryos were collected from superovulated female BDF1 mice. The collected embryos were randomly divided into three groups. One group was maintained as fresh controls (n=42), one was frozen by slow freezing (n=43), and one was cooled by vitrification (n=43). After thawing or cooling, survival rates, development to blastocyst, and cell numbers and inner cell mass (ICM) cell numbers of blastocysts were compared with those of the control group. The expressions of eight genes (Rbm3, Birc5, Sod1, Sod2, Cirbp, Caspase3, Trp53, Hsp70.1) were examined by real time-quantitative polymerase chain reaction in the fresh and frozen-thawed embryos.ResultsThere were no significant differences in the slow freezing and vitrification groups' survival rate after thawing (88.4% vs. 88.4%), development to blastocyst (100% vs. 97.4%), cell numbers (107.0±21.0 vs. 115.0±19.7), or ICM cell numbers of blastocysts (11.3±5.2 vs. 11.1±3.7). Cell numbers of blastocysts were significantly (p<0.05) lower in the frozen-thawed embryos than the fresh embryos. There were no significant differences in the slow freezing and the vitrification groups' expressions of the eight genes. The expressions of CirbP and Hsp70.1 were higher in the frozen-thawed embryos than in the fresh embryos but there were no significant differences.ConclusionThese results suggest that there were no significant differences between embryos that underwent slow freezing and vitrification.

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“…Recently many clinics have begun to vitrify embryos as an alternative to freezing, mainly because of the higher rates of survival obtained (Rama Raju et al, 2005; Balaban et al, 2008). However the clinical superiority of vitrification over freezing has not been demonstrated in RCTs (Kolibianakis et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

A pragmatic RCT of conventional versus increased concentration sucrose in freezing and thawing solutions for human embryos

et al. 2011

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BACKGROUNDIntact frozen-thawed embryos have a greater potential than damaged embryos to establish successful pregnancies. This study aimed to determine whether elevated concentrations of sucrose during freezing would increase the proportion of patients with ≥50% of embryos intact after thawing (primary outcome), and improve clinical outcome.METHODSIn a two arm, parallel group, pragmatic trial, IVF/ICSI couples were randomized prospectively to have their supernumerary embryos frozen in a medium containing 0.1 M sucrose (control; n = 99) or 0.3 M sucrose (intervention; n = 102).RESULTSMore control (74/99) than intervention (63/102) couples had at least one embryo thawed (P = 0.07). Significantly more (P = 0.005) intervention (53/63) than control (45/74) couples had ≥50% of embryos intact. Freezing in a medium containing 0.3 M sucrose increased by 3.4-fold [95% confidence interval (CI) (1.45, 7.82)] the likelihood of a couple having ≥50% of their embryos intact. In the fresh cycle, live birth rate per transfer was similar in the control (35/95) and intervention (36/93) groups (P = 0.91). More control (19/63) than intervention (9/59) couples had a live birth after frozen embryo transfer (P = 0.08). When fresh and frozen cycles were combined, fewer intervention (n = 102) than control (n = 99) couples had at least one live birth (42 versus 53%). The difference in cumulative live birth rate was not significant [hazard ratio = 0.75, 95% CI (0.49, 1.13); P = 0.17].CONCLUSIONSIncreasing the concentration of sucrose in the freezing medium improves embryo survival, but this is not reflected by increased cumulative birth rates.Clinical Trials Registration number: ISRCTN93314892.

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Vitrification of human 8-cell embryos, a modified protocol for better pregnancy rates

Cited by 109 publications

References 23 publications

A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos

A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos

In vitrodevelopment and gene expression of frozen-thawed 8-cell stage mouse embryos following slow freezing or vitrification

A pragmatic RCT of conventional versus increased concentration sucrose in freezing and thawing solutions for human embryos

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