The worldwide frozen embryo reservoir: methodologies to achieve optimal results

Capalbo, Antonio; Rienzi, Laura; Buccheri, Matteo; Maggiulli, Roberta; Sapienza, Fabio; Romano, Stefania; Colamaria, Silvia; Iussig, Benedetta; Giuliani, Maddalena; Palagiano, Antonio; Ubaldi, Filippo Maria

doi:10.1111/j.1749-6632.2010.05931.x

Cited by 15 publications

(5 citation statements)

References 48 publications

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“…1 Researchers were initially concerned with the effects of cryopreservation or cryostorage on the survival, development and molecular biology of the frozen embryo after thawing. 2–4 However, recent reports have demonstrated that cryostorage does not adversely affect post-thaw survival or pregnancy outcome during in vitro fertilization (IVF) or oocyte donation in patients.…”

Section: Introductionmentioning

confidence: 99%

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

Pruksananonda

Rungsiwiwut

Numchaisrika

et al. 2012

BioResearch Open Access

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Human embryonic stem (hES) cells are considered to be a potential source for the therapy of human diseases, drug screening, and the study of developmental biology. In the present study, we successfully derived hES cell lines from blastocysts developed from frozen and fresh embryos. Seventeen- to eighteen-year-old frozen embryos were thawed, cultured to the blastocyst stage, and induced to form hES cells using human foreskin fibroblasts. The Chula2.hES cell line and the Chula4.hES and Chula5.hES cell lines were derived from blastocysts developed from frozen and fresh embryos, respectively. The cell lines expressed pluripotent markers, including alkaline phosphatase (AP), Oct3/4, stage-specific embryonic antigen (SSEA)-4, and tumor recognition antigen (TRA)-1-60 and TRA-1-81 as detected with immunocytochemistry. The real-time polymerase chain reaction (RT-PCR) results showed that the cell lines expressed pluripotent genes, including OCT3/4, SOX2, NANOG, UTF, LIN28, REX1, NODAL, and E-Cadherin. In addition, the telomerase activities of the cell lines were higher than in the fibroblast cells. Moreover, the cell lines differentiated into all three germ layers both in vitro and in vivo. The cell lines had distinct identities, as revealed with DNA fingerprinting, and maintained their normal karyotype after a long-term culture. This study is the first to report the successful derivation of hES cell lines in Thailand and that frozen embryos maintained their pluripotency similar to fresh embryos, as shown by the success of hES cell derivation, even after years of cryopreservation. Therefore, embryos from prolonged cryopreservation could be an alternative source for embryonic stem cell research.

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Section: Introductionmentioning

confidence: 99%

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

Pruksananonda

Rungsiwiwut

Numchaisrika

et al. 2012

BioResearch Open Access

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“…Embryo cryopreservation has become an established procedure in the field of reproductive medicine [12, 30–32] accounting for a cumulative 28% of delivery rate following IVF programs at least in Europe [4, 10, 33]. Slow-freezing was the first technique of cryopreservation employed in IVF laboratories, but, in the last years, it has been progressively replaced by the method of vitrification of the basis of a quantity of literature recently reviewed reporting higher cryosurvival rate in both cleavage- and blastocyst-stage embyos [23].…”

Section: Discussionmentioning

confidence: 99%

A monocentric analysis of the efficacy of extracellular cryoprotectants in unfrozen solutions for cleavage stage embryos

Capodanno

Daolio

Feo

et al. 2019

Reprod Biol Endocrinol

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BackgroundIn the absence of international guidelines indicating the usage of vitrification rather than slow-freezing, the study aim was to analyze a large cohort of slow-frozen/thawed embryos to produce a rationale supporting the standardization of IVF cryopreservation policy.MethodsThis retrospective analysis included 4779 cleavage stage embryos cryopreserved by slow-freezing/thawing from September 2009 to April 2017 at a single Center. Biological and clinical outcomes of three different commercial kits adopted sequentially, i.e. Vitrolife Cleave Kit® from Vitrolife (kit 1) vs. K-SICS-5000 Kit® and K-SITS-5000 Kit® from Cook Medical (kit 2) and Freeze/Thaw 1™ Kit® from Vitrolife (kit 3) were collected and compared in the light of cryoprotectants composition.ResultsKit 3 compared to kit 1 and kit 2 showed significantly (P < 0.001) higher embryo survival (79.9% vs. 75.6 and 68.1%, respectively) and frozen embryo replacement (91.5% vs. 86.5 and 83.3%, respectively) rates, and significantly (P < 0.001) lower blastomere degeneration rate (41.5% vs. 43.6 and 52.4%, respectively). No significant difference for clinical outcomes was observed among kits. Only a slight positive trend was observed for kit 3 vs. kit 1 and kit 2 on delivery rate per thawing cycle (7.12% vs. 4.19 and 4.51%, respectively; P < 0.058) and live birth rate (3.07% vs. 2.59 and 1.93%, respectively, P < 0.069). Thawing solutions of kit 3 were similar to those of any warming protocol.ConclusionsA defined concentration of extracellular cryoprotectants in thawing/warming solutions had a beneficial effect on the embryo cryosurvival rate. Results could provide the rationale for the adoption of a single standardized warming protocol.

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“…It is known that zona pellucida and mucin coat are essential for rabbit embryo development and implantation [28] and damage to zona pellucida is common phenomenon when embryos are cryopreserved in normal straws [44,30,58,20]. On the other hand, cryopreservation by ultra-rapid vitrification utilizing the MEV method and accelerated cooling/warming rates might avoid such damage [8]. Likewise, high recovery rates of the zona pellucida-intact pronuclear zygotes might also be attributed to the composition of vitrification media.…”

Section: Discussionmentioning

confidence: 99%

State of actin cytoskeleton and development of slow-frozen and vitrified rabbit pronuclear zygotes

et al. 2016

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This study was focused on the effect of cryopreservation on the state of actin cytoskeleton and development of rabbit pronuclear zygotes. Zygotes were collected from superovulated females and immediately used for 1) slow-freezing in a solution containing 1.5 M 1,2-propanediol and 0.2 M sucrose, or 2) vitrification in a solution containing 42.0% (v/v) of ethylene glycol, 18.0% (w/v) of dextran and 0.3 M sucrose as cryoprotectants. After thawing or warming, respectively, zygotes were evaluated for 1) actin distribution, 2) in vitro or 3) in vivo development to blastocyst. Comparing actin filaments distribution, a significantly higher number of vitrified zygotes with actin distributed in cell border was observed (55 ± 7.7 vs. 74 ± 6.1% for slow-frozen vs. vitrified, respectively). After 24 and 72 h of in vitro development, significant differences in the cleavage and morula rate among the groups were observed (9 ± 2.4 and 3 ± 1.3 vs. 44 ± 3.0 and 28 ± 2.7% for slow-frozen vs. vitrified, respectively). None of the slow-frozen zygotes reached the blastocyst stage, in contrast to the vitrified counterparts (11 ± 1.9%). Under in vivo culture conditions, a significant difference in blastocyst rate was observed between vitrified and fresh embryos (6 ± 1.5 vs. 35 ± 4.4% respectively). Our results showed that alterations in actin cytoskeleton and deteriorated development are more evident in slow-frozen than vitrified pronuclear zygotes. Vitrification method seems to be a more effective option for rabbit zygotes cryopreservation, although pronuclear zygotes manipulation per se resulted in a notable decrease in embryo development.

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The worldwide frozen embryo reservoir: methodologies to achieve optimal results

Cited by 15 publications

References 48 publications

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

A monocentric analysis of the efficacy of extracellular cryoprotectants in unfrozen solutions for cleavage stage embryos

State of actin cytoskeleton and development of slow-frozen and vitrified rabbit pronuclear zygotes

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