Cryopreservation of human embryos using ethylene glycol in controlled slow freezing

Chi, Hee-Jun; Koo, Jung-Jin; Kim, Moon-Young; Joo, Jin-Young; Chang, Sang-Sik; Chung, Kil-Saeng

doi:10.1093/humrep/17.8.2146

Cited by 69 publications

(30 citation statements)

References 32 publications

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“…Ethylene Glycol has proven to be a stable cryoprotectant, with less toxicity and high permeation ability suggested that Ethylene Glycol diffuses into and leaves the embryos very rapidly due to its low molecular weight, hence embryos do not undergo osmotic shock during freezing and thawing procedures. 21,22 The embryo survival rate of 96.95% with 94.17% having excellent morphology in our study, further supports this concept. Mukaida et al (1998) reported a survival rate of 81% with EG based media.…”

Section: Discussionsupporting

confidence: 82%

Comparison of vitrification and slow freezing for cryopreservation of cleavage stage embryos (Day 3) and its impact on clinical outcome

et al. 2015

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Section: Discussionsupporting

confidence: 82%

Comparison of vitrification and slow freezing for cryopreservation of cleavage stage embryos (Day 3) and its impact on clinical outcome

et al. 2015

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“…It appears that the damage to the follicles observed using this cryoprotectant was caused by a toxic effect of the EG and not to the cryopreservation process. In previous studies, EG was nontoxic to human [16] and bovine [17] and [18] embryos. Moreover, good results were observed when EG was used to cryopreserve murine ovarian tissue, with 88% of follicles remaining morphologically normal after freezing/thawing [4].…”

Section: Discussionmentioning

confidence: 77%

Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bos indicus) ovarian tissue

et al. 2004

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Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bos indicus) ovarian tissueCarolina M Lucci Mirella A Kacinskis Luiz Henrique R Lopes Rodolfo Rumpf Sônia N Báoa AbstractCryopreservation of ovarian tissue is a new and promising technique for germ-line storage. The objective of this study was to evaluate the effect of four cryoprotectants (at two concentrations each) on the preservation of zebu bovine preantral follicles after ovarian cryostorage. Strips of ovarian cortex were cryopreserved using glycerol (GLY; 10 or 20%), ethylene glycol (EG), propanediol (PROH) or dimethylsulphoxide (DMSO; 1.5 or 3 M). In addition, a toxicity test was performed for each cryoprotectant by exposing the ovarian tissue to them without freezing. Tissues were analyzed by histology and transmission electron microscopy. Ovarian tissue frozen in either concentration of DMSO or PROH or in 10% GLY retained a higher percentage of morphologically normal follicles (73-88%) than tissue frozen in 20% GLY or in either concentration of EG (16-52%). In the toxicity test, exposure of tissues to DMSO, PROH or GLY resulted in higher percentages of normal follicles (80-97%) than exposure to EG (49%). Electron microscopy revealed damage to the ultrastructure of follicles frozen in 10% GLY, while follicles cryopreserved in DMSO and PROH at either concentration exhibited normal ultrastructure. In conclusion, DMSO and PROH were the most effective cryoprotectants for zebu ovarian tissue, preserving the structural integrity of somatic and reproductive cells within the ovary.

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“…Furthermore, ethylene g l y c o l ( E G ) h as b e e n u s e d w i d e l y f o r t h e vitrification of human oocytes [10,25] and embryos [26,27,30,31,[36][37][38] due to its low molecular weight, high permeation ability and low toxicity. Moreover, it has been reported that cryoprotectant mixtures may have some advantages over solutions containing only one permeable cryoprotectant [16].…”

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confidence: 99%

High Survival Rate of Bovine Oocytes Matured In Vitro Following Vitrification

Chian

Kuwayama

Tan

et al. 2004

Journal of Reproduction and Development

166

100

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Abstract. Improving pregnancy rates associated with the use of cryopreserved human oocytes would be an important advance in human assisted reproductive technology (ART). Vitrification allows glasslike solidification of a solution without ice crystal formation in the living cells. We have attempted to improve the survival rates of oocytes by a vitrification technique using bovine models. In vitro matured oocytes with or without cumulus cells were vitrified with either 15.0% (v/v) ethylene glycol (EG) + 15% (v/v) dimethylsulfoxide (DMSO) + 0.5 M sucrose or 15% (v/v) EG + 15% (v/v) 1,2-propanediol (PROH) + 0.5 M sucrose, using 'Cryotop' or 'thin plastic sticker', respectively. The oocyte survival rates after vitrifying-warming, and the capacity for fertilization and embryonic development were examined in vitro. The rate of embryonic development to blastocyst was significantly higher (P<0.05) in the oocytes vitrified with 15% (v/v) EG + 15% (v/v) PROH + 0.5 M sucrose than in the oocytes vitrified with 15% (v/v) EG + 15% (v/v) DMSO + 0.5 M sucrose (7.4% ± 4.1 vs. 1.7% ± 3.0, respectively). Oocytes vitrified without cumulus cells had a higher survival rate after thawing and a superior embryonic developmental capacity compared with oocytes vitrified with cumulus cells. Prolonged pre-incubation time after thawing adversely affected the rates of embryonic cleavage and development. These results indicate that in vitro matured bovine oocytes can be vitrified successfully with the mixture of the cryoprotectants, EG + PROH, the absence of cumulus cells for vitrification does not affect oocyte survival rate after warming, and vitrified and warmed oocytes do not require pre-incubation before in vitro fertilization.

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Cryopreservation of human embryos using ethylene glycol in controlled slow freezing

Abstract: Ethylene glycol may be a good substitute for PROH to cryopreserve human embryos using a slow-freezing protocol.

Cited by 69 publications

References 32 publications

Comparison of vitrification and slow freezing for cryopreservation of cleavage stage embryos (Day 3) and its impact on clinical outcome

Comparison of vitrification and slow freezing for cryopreservation of cleavage stage embryos (Day 3) and its impact on clinical outcome

Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bos indicus) ovarian tissue

High Survival Rate of Bovine Oocytes Matured In Vitro Following Vitrification

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