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Hum. Reprod. Advance Access published online on January 21, 2005
Human Reproduction, doi:10.1093/humrep/deh742
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Human Reproduction © European Society of Human Reproduction and Embryology 2005; all rights reserved
Received March 8, 2004
Revised December 7, 2004
Accepted December 13, 2004

Article

Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures

S.J. Paynter 1*, A. Borini 2, V. Bianchi 2, L. De Santis 3, C. Flamigni 4, and G. Coticchio 2

1 Department of Obstetrics and Gynaecology, Wales College of Medicine, Cardiff University, Cardiff CF14 4XN, UK
2 Tecnobios Procreazione, Via Dante 15,, Italy
3 Department of Obstetrics and Gynaecology, Vita-Salute University, H S. Raffaele, 20132 Milan, Italy
4 University of Bologna, 40125 Bologna, Italy and

* To whom correspondence should be addressed.
S.J. Paynter, E-mail: Paynter{at}cardiff.ac.uk


  Abstract

BACKGROUND: Despite the recent increase in pregnancies from cryopreserved human oocytes, success in terms of births per thawed oocyte is still poor. Modifications to cryopreservation protocols have not been based on measurement of the osmotic response of oocytes, and methodologies are often poorly described or protocols not strictly adhered to, inevitably resulting in variability. METHODS: Volume change of mature human oocytes was measured on exposure to cryoprotectant. Oocytes were exposed to either 0.75 mol/l propane-1,2-diol (PrOH) for 10 min; 1.5 mol/l PrOH for 10 min, having been exposed to 0.75 mol/l PrOH for 7.5 min; or 1.5 mol/l PrOH plus 0.2 or 0.3 mol/l sucrose for 10 min, having been exposed to 1.5 mol/l PrOH for 10 min. RESULTS: On exposure to PrOH alone, oocytes shrank and then re-expanded, having reached 75 and 84% of their starting volume in 0.75 and 1.5 mol/l, respectively. Oocytes shrank continuously in PrOH plus sucrose, reaching 67 or 55% of their initial volume in 0.2 or 0.3 mol/l sucrose, respectively. CONCLUSIONS: To improve consistency following cryopreservation, protocols must be strictly adhered to; small changes in duration of exposure to cryoprotectant can result in drastic changes in cellular hydration and thus the fate of the cell during freezing/thawing.

Keywords: cryopreservation; cryoprotectant; oocyte; permeability; propane-1,2-diol.
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