Vitrification of Mature Mouse Oocytes: Improved Results Following Addition of Polyethylene Glycol to a Dimethyl Sulfoxide Solution

Authors: O'Neil L.; Paynter S.J.; Fuller B.J.; Shaw R.W.

Source: Cryobiology, Volume 34, Number 3, May 1997 , pp. 295-301(7)

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Abstract:

Oocytes have been successfully cryopreserved using rapid and slow freezing procedures; however, variability in the success of replicates has limited its practical application. We have evaluated the potentially beneficial effects of adding 1 mg/ml of the polymer polyethylene glycol (PEG) ( M r 8000) to a 6 M dimethyl sulfoxide (Me 2 SO) vitrification solution. Stepwise addition of cryoprotectant, either with or without PEG, was performed at room temperature (19-21#&176;C). Oocytes were then loaded in plastic insemination straws and held in liquid nitrogen vapour at -140#&176;C for 3 min prior to storage in liquid nitrogen. Oocytes were warmed rapidly to room temperature and removal of cryoprotective agent was effected in the presence of 1 M sucrose solution. Viability was assessed by in vitro fertilization. Oocytes cryopreserved after exposure to 6 M Me 2 SO in the absence of PEG showed 60% normality, 80% fertilization, and 55% development to blastocyst, median of 11 replicate experiments (191 oocytes). Individual replicates yielded highly variable survival which ranged from 0 to 100%. The addition of PEG significantly improved oocyte normality to 95% (range, 76-100%; median of 9 replicate experiments, 301 oocytes). Rates of fertilization (91%; 60-100%) and development to blastocyst (73%; 67-92%) were also improved. The addition of 1 mg/ml PEG to a 6 M Me 2 SO solution resulted in greatly improved viability of oocytes following cryopreservation and vastly reduced the variability seen with the Me 2 SO solution alone.

Language: English

Document Type: Research article

Affiliations: Department of Obstetrics and Gynaecology, University of Wales College of Medicine, Heath Park, Cardiff, Wales, CF4 4XN, United Kingdom

Publication date: 1997-05-01

Vitrification of Mature Mouse Oocytes: Improved Results Following Addition of Polyethylene Glycol to a Dimethyl Sulfoxide Solution

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