Cryobiological Preservation of
            <i>Drosophila</i>
            Embryos

Mazur, Péter; Cole, Kenneth W.; Hall, Jerry W.; Schreuders, Paul; Mahowald, Anthony P.

doi:10.1126/science.1470915

Cited by 159 publications

(81 citation statements)

References 16 publications

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“…A key aim when developing the OVS was to maximise the rate of cooling, minimising the extent of ice crystal formation without the need for high concentrations of cryoprotectant [20]. This reflects a general focus on cooling rate in the development of vitrification for mammalian embryos [26].…”

Section: Discussionmentioning

confidence: 99%

“…Cryopreservation by vitrification occurs by glass formation both inside and outside the sample after dehydration. Two main challenges to refining the vitrification technique have been identified: i) high concentrations of cryoprotectant are required, and these may be toxic to oocytes, embryos, and ovaries; and ii) intracellular ice and chilling may occur if the applied cooling or warming rates are insufficient [17,20,30]. To overcome these problems, several methods have been developed.…”

Section: Introductionmentioning

confidence: 99%

“…The most logical way to increase the cooling rate is to use the smallest possible volume of cryoprotectant medium to surround the sample and then expose it directly to liquid nitrogen without any thermo-insulation: this is the open vitrification system (OVS). This idea was initially proposed for freezing Drosophila embryos (Mazur et al 1992). Animal data [11,15,22,31] and recent analyses of clinical reports [18,19] have revealed the benefits of vitrification such as low rates of cellular damage.…”

Section: Introductionmentioning

confidence: 99%

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A closed system supports the developmental competence of human embryos after vitrification

Hashimoto

Amo

Hama

et al. 2013

J Assist Reprod Genet

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Purpose Closed-system vitrification may enable the risk of contamination to be minimised. We performed three studies to compare the developmental competence of human embryos vitrified using either a closed vitrification system (CVS; Rapid-i®) or an open vitrification system (OVS; Cryo-top®). Methods The first study was performed in vitro using 66 zygotes previously vitrified at pronuclear stage. These were warmed and randomised 1:1 to revitrification using either the OVS or the CVS. After re-warming, embryo development and blastocyst cell number were assessed. For the second study, also performed in vitro, 60 vitrified-warmed blastocysts were randomised 1:1:1 into three groups (OVS or CVS revitrification, or no revitrification). The proportion of dead cells was assessed by staining. The third study was performed in vivo, using 263 high-grade blastocysts randomly assigned to vitrification using either the CVS (n=100) or the OVS (n=163). After warming, single blastocyst transfer was performed. Results There were no differences between the CVS and the OVS in survival rate (100 % vs. 97 %), blastulation rate (96 h: 50 % vs. 50 %; 120 h: 68 % vs. 56 %), proportion of good blastocysts (96 h: 32 % vs. 22 %, 120 h: 47 % vs. 41 %), or mean number of cells (137 vs. 138). The proportion of dead cells in blastocysts re-vitrified by CVS (31 %) was similar to that for OVS (38 %) and non-revitrification (32 %). In vivo, the implantation rate for blastocysts vitrified using the CVS (54 %) was similar to that with the OVS (53 %).Conclusion Our studies consistently indicate that human embryos may be vitrified using a CVS without impairment of developmental competence.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A closed system supports the developmental competence of human embryos after vitrification

Hashimoto

Amo

Hama

et al. 2013

J Assist Reprod Genet

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“…Slower cooling would increase the risk of ice crystal formation in the cells [4,6]. The average rate of cooling is about −1220°C/min in the Rapid-i device [35], whereas that of the Cryotop (open vitrification) device is −23000°C/min [15].…”

Section: Discussionmentioning

confidence: 99%

“…This procedure is known as an open vitrification system. This idea was initially proposed for freezing Drosophila embryos [6], and drastic improvement in viability has since been shown in both animal studies [7][8][9][10] and clinical reports [11,12]. However, there are some potential drawbacks of the open vitrification system, such as the sterility of liquid nitrogen and the risk of cross-contamination during long-term storage [13,14].…”

Section: Introductionmentioning

confidence: 99%

Neonatal outcomes after the implantation of human embryos vitrified using a closed-system device

Iwahata

Hashimoto

Inoue³

et al. 2015

J Assist Reprod Genet

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Purpose Closed vitrification poses a risk of adversely affecting embryo development, while it may minimize the risk of contamination. We assessed the effects of closed-system human embryo vitrification on fetal development after implantation, neonatal outcome, and clinical safety. Methods This was a retrospective cohort study conducted at a private fertility clinic. A total of 875 vitrified-warmed blastocysts that were single-transferred under hormone-replacement cycles between November 2011 and December 2013 were randomly divided into two groups (closed vitrification, n 313; open vitrification, n 562) after receiving the patients' consent forms. Developmental competence after implantation, including gestational age, birth weight, sex, Apgar score, and anomalies of newborns, after the transfer of blastocysts vitrified by closing vitrification was compared with that obtained in the case of open vitrification. Results There were no significant differences between the use of closed and open vitrification systems in embryo development after implantation, gestational age, birth weight, sex ratio, Apgar score, and congenital anomalies of newborns. Conclusion Human embryos can be vitrified using a closed vitrification system without impairment of neonatal development.

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Water distribution and permeability of zebrafish embryos,Brachydanio rerio

Hagedorn¹,

Kleinhans²,

Freitas³

et al. 1997

J. Exp. Zool.

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Teleost embryos have not been successfully cryopreserved. To formulate successful cryopreservation protocols, the distribution and cellular permeability to water must be understood. In this paper, the zebrafish (Brachydanio rerio) was used as a model for basic studies of the distribution to permeability to water. These embryos are a complex multi-compartmental system composed of two membrane-limited compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells (each surrounded by a plasma membrane). Due to the complexity of this system, a variety of techniques, including magnetic resonance microscopy and electron spin resonance, was used to measure the water in these compartments. Cellular water was distributed unequally in each compartment. At the 6-somite stage, the percent water (VA'^) was distributed as follows: total in embryo = 74%, total in yolk = 42%, and total in blastoderm = 82%. A one-compartment model was used to analyze kinetic, osmotic shrinkage data and determine a phenomenological water permeability parameter, Lp, assuming intracellular isosmotic compartments of either 40 or 300 mosm. This analysis revealed that the membrane permeability changed (P< 0.05) during development. During the 75% epiboly to 3-somite stage, the mean membrane permeability remained constant (Lp = 0.022 ± 0.002 |xm x min'^atm"^ [mean ± S.E.M.] assuming isosmotic is 40 mosm or Lp = 0.049 ± 0.008 |im x min'^atm"^ assuming isosmotic is 300 mosm). However, at the 6-somite stage, Lp increased twofold (Lp = 0.040 ± 0.004 |j,m x min'^atm"^ assuming isosmotic is 40 mosm or Lp = 0.100 ± 0.017 |J,m x min'^atm"^ assuming isosmotic is 300 mosm). Therefore, the low permeability of the zebrafish embryo coupled with its large size (and consequent low area to volume ratio) led to a very slow osmotic response that should be considered before formulating cryopreservation protocols.

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Cryobiological Preservation of Drosophila Embryos

Cited by 159 publications

References 16 publications

A closed system supports the developmental competence of human embryos after vitrification

A closed system supports the developmental competence of human embryos after vitrification

Neonatal outcomes after the implantation of human embryos vitrified using a closed-system device

Water distribution and permeability of zebrafish embryos,Brachydanio rerio

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