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Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification

Published online by Cambridge University Press:  07 December 2018

Jaqueline Sudiman*
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Alice Lee
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Kheng Ling Ong
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Wu Zi Yuan
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Sarah Jansen
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Peter Temple-Smith
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Mulyoto Pangestu
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Sally Catt
Obstetrics and Gynaecology Department, Monash Medical Centre, 246 Clayton Road, Victoria, Australia
Address for correspondence: Jaqueline Sudiman. Monash Institute of Medical Research, Obstetrics and Gynaecology Department, Udayana University, Denpasar, Bali, Indonesia. Tel: +62 82283387245. E-mail:


Mouse and lamb oocytes were vitrified with, or exposed to, different cryoprotectants and evaluated for their effects on their survival and developmental competence after in vitro fertilization (IVF) and activation treatments. Control oocytes remained untreated, whilst the remainder were exposed to three different combinations of vitrification solutions [dimethyl sulfoxide (DMSO) + ethylene glycol (EG), EG only, or propanediol (PROH) + EG] and either vitrified or left unfrozen (exposed groups). Oocytes in the control and vitrified groups underwent IVF and developmental competence was assessed to the blastocyst stage. In lambs, survival rate in vitrified oocytes was significantly lower than for oocytes in the exposed groups (P <0.05). Blastocyst development was low in vitrified oocytes compared with controls (<6% vs 38.9%, P <0.01). Parthenogenetic activation was more prevalent in vitrified lamb oocytes compared with controls (P <0.05). No evidence of zona pellucida hardening or cortical granule exocytosis could account for reduced fertilization rates in vitrified lamb oocytes. Mouse oocytes demonstrated a completely different response to lamb oocytes, with survival and parthenogenetic activation rates unaffected by the vitrification process. Treatment of mouse oocytes with DMSO + EG yielded significantly higher survival and cleavage rates than treatment with PROH + EG (87.8% and 51.7% vs 32.7% and 16.7% respectively, P <0.01), however cleavage rate for vitrified oocytes remained lower than for the controls (51.7% vs 91.7%, P <0.01) as did mean blastocyst cell number (33 ± 3.1 vs 42 ± 1.5, P <0.05). From this study, it is clear that lamb and mouse show different tolerances to cryoprotectants commonly used in vitrification procedures, and careful selection and testing of species-compatible cryoprotectants is required when vitrifying oocytes to optimize survival and embryo development.

Research Article
© Cambridge University Press 2018 

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Affiliated to the Obstetrics and Gynaecology Department, Udayana University, Denpasar, Bali, Indonesia.


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