Slow and ultrarapid cryopreservation of biopsied mouse blastocysts and its effect on DNA integrity index

Kader, A.; Falcone, Tommaso; Sharma, Rakesh; Mangrola, Devna; Agarwal, Ashok

doi:10.1007/s10815-010-9441-4

Cited by 11 publications

(13 citation statements)

References 29 publications

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“…In this regard, we assessed the integrity of the vitrifiedwarmed blastocysts through determining the total cell number and the apoptosis rate. Percentages of apoptotic cells were significantly higher in vitrified than in fresh blastocysts, in agreement with previous studies (Kader et al 2009(Kader et al , 2010Castillo-Martín et al 2013, 2014. In contrast, there were no differences in cell number, which is also in accordance with previous works (Cuello et al 2007;Du et al 2007;Castillo-Martín et al 2014).…”

Section: Discussionsupporting

confidence: 93%

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Morató

Castillo‐Martín

Yeste

et al. 2016

Reprod. Fertil. Dev.

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The aim of our study was to assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity before vitrification, using the pig as a model. For this purpose we analysed the cryoresistance and apoptosis rate of blastocysts at different stages of development as derived on Day 5 and 6 of in vitro culture. Blastocysts were subsequently vitrified, warmed and cultured for 24h. Re-expansion rates were recorded at 3 and 24h and total cell number and apoptotic cells were determined at 24h. Day-6 blastocysts showed the highest rates of survival after warming, which indicates higher quality compared with Day-5 blastocysts. Higher re-expansion rates were observed for expanded blastocysts and those in the process of hatching when compared with early blastocysts. Total cell number and apoptotic cells were affected by blastocyst stage, vitrification-warming procedures and length of in vitro culture, as expanding and hatching-hatched blastocysts from Day 6 presented higher percentages of apoptotic cells than fresh blastocysts and blastocysts vitrified at Day 5. Our findings suggest that the cryotop vitrification method is useful for the cryopreservation of porcine blastocysts presenting a high degree of expansion, particularly when vitrification is performed after 6 days of in vitro culture. Furthermore, these results show that faster embryo development underlies higher blastocyst cryotolerance and provide evidence that blastocoel cavity expansion before vitrification is a reliable index of in vitro-produced embryo quality and developmental potential.

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Section: Discussionsupporting

confidence: 93%

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Morató

Castillo‐Martín

Yeste

et al. 2016

Reprod. Fertil. Dev.

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“…These results are similar to those of Kader et al. () who reported that both slow freezing and vitrification of biopsied mouse blastocysts induced apoptosis to a similar extent. The number of apoptotic nuclei and the apoptotic index was significantly (p < 0.05) lower for the control non‐cryopreserved cloned embryos compared with those cryopreserved by slow freezing or vitrification.…”

Section: Discussionsupporting

confidence: 92%

Cryopreservation of Zona‐Free Cloned Buffalo (Bubalus Bubalis) Embryos: Slow Freezing vs Open‐Pulled Straw Vitrification

Sirisha

Selokar

Saini

et al. 2013

Reprod Domestic Animals

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This study was carried out to compare the post-thaw cryosurvival rate and the level of apoptosis in vitro produced zona-free cloned buffalo blastocysts subjected to slow freezing or vitrification in open-pulled straws (OPS). Zona-free cloned embryos produced by handmade cloning were divided into two groups and were cryopreserved either by slow freezing or by vitrification in OPS. Cryosurvival of blastocysts was determined by their re-expansion rate following post-thaw culture for 22-24 h. The post-thaw re-expansion rate was significantly (p < 0.05) higher following vitrification in OPS (71.2 ± 2.3%) compared with that after slow freezing (41.6 ± 4.8%). For examining embryo quality, the level of apoptosis in day 8 frozen-thawed blastocysts was determined by TUNEL staining. The total cell number was not significantly different among the control non-cryopreserved cloned embryos (422.6 ± 67.8) and those cryopreserved by slow freezing (376.4 ± 29.3) or vitrification in OPS (422.8 ± 36.2). However, the apoptotic index, which was similar for embryos subjected to slow freezing (14.8 ± 2.0) or OPS vitrification (13.3 ± 1.8), was significantly (p < 0.05) higher than that for the control non-cryopreserved cloned embryos (3.4 ± 0.6). In conclusion, the results of this study demonstrate that vitrification in OPS is better than slow freezing for the cryopreservation of zona-free cloned buffalo blastocysts because it offers a much higher cryosurvival rate.

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“…Information about the effects of CP on chromatin integrity are incomplete and contradictory. Kader et al [ 19 ] investigated the chromatin integrity index in fresh, vitrified and slow-frozen blastocysts and found significant decrease in vitrified embryos. However, Li et al [ 20 ] found higher DNA integrity in vitrified human and mouse blastocysts than in the slow-frozen ones and similar to fresh blastocysts.…”

Section: Introductionmentioning

confidence: 99%

Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification

Somoskői

Martino

Cardone

et al. 2015

Reprod Biol Endocrinol

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BackgroundThe ability to cryopreserve mammalian embryos has become an integral part of assisted reproduction, both in human and veterinary medicine. Despite differences in the size and physiological characteristics of embryos from various species, the embryos have been frozen by either of two procedures: slow freezing or vitrification. The aim of our study was to compare the effect of slow freezing and vitrification to the chromatin structure, energy status and reactive oxygen species production of mouse morulae and blastocysts.MethodsMouse morulae and blastocysts were randomly allocated into vitrification, slow freezing and control groups. For slow freezing, Dulbecco phosphate buffered saline based 10% glicerol solution was used. For vitrification, G-MOPS™ based solution supplemented with 16% ethylene glycol, 16% propylene glycol, Ficoll (10 mg/ml) and sucrose (0.65 mol/l) was used. After warming, the chromatin integrity, mitochondrial distribution pattern and energy/oxidative status were compared among groups.ResultsCryopreservation affected chromatin integrity at a greater extent at the morula than the blastocyst stage. Chromatin damage induced by slow freezing was more relevant compared to vitrification. Slow freezing and vitrification similarly affected mitochondrial distribution pattern. Greater damage was observed at the morula stage and it was associated with embryo grade. Cryopreservation altered the quantitative bioenergy/redox parameters at a greater extent in the morulae than in the blastocysts. Effects induced by slow freezing were not related to embryo grade or mitochondrial pattern, as affected embryos were of all grades and with both mitochondrial patterns. However, effects induced by vitrification were related to mitochondrial pattern, as only embryos with homogeneous mitochondrial pattern in small aggregates had reduced energy status.ConclusionsThis study shows for the first time the joint assessment of chromatin damage and mitochondrial energy/redox potential in fresh and frozen mouse embryos at the morula and blastocyst stage, allowing the comparison of the effects of the two most commonly used cryopreservation procedures.

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Slow and ultrarapid cryopreservation of biopsied mouse blastocysts and its effect on DNA integrity index

Abstract: Slow freezing and vitrification are equal options for preservation of biopsied blastocysts as regards the DNA integrity index (DII). Biopsied blastocysts better tolerate vitrification than intact expanded blastocysts.

Cited by 11 publications

References 29 publications

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Cryopreservation of Zona‐Free Cloned Buffalo (Bubalus Bubalis) Embryos: Slow Freezing vs Open‐Pulled Straw Vitrification

Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification

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