Effect of the cryoprotectant concentration on the in vitro embryo development and cell proliferation of OPS-vitrified porcine blastocysts.

Abstract:

:Our objective was to study the effect of the concentration of ethylene glycol (EG) and dimethyl sulfoxide (Me2SO) during vitrification on the development of porcine blastocysts. Vitrification was performed with 0.4 M sucrose and either a Me2SO and EG mixture (15%, 16% and 17% v/v of each) or EG alone (40% v/v), using superfine open pulled straws. Fresh and vitrified blastocysts were cultured for 48 h and the survival and hatching rates were evaluated. Some vitrified and fresh embryos were processed for Hoechst 33342 staining and proliferation cell nuclear antigen (PCNA) inmunolocalization to determine the proliferation index. The survival rate was similar for fresh and vitrified blastocysts, except for blastocysts vitrified using 15% of cryoprotectants, which displayed lower (P < 0.05) survival than fresh blastocysts. Vitrified and fresh blastocysts had a similar cell proliferation index (range: 75.8+/-3.2 to 83.7+/-3). When only hatched blastocysts among groups were compared, the proliferation rate decreased (P < 0.05) after vitrification with 17% of EG-Me2SO. In conclusion, the concentration of EG-Me2SO could be decreased to 16% in the vitrification medium with no reduction of the in vitro developmental ability of the blastocysts. In addition, a 40% EG-based medium can be used for vitrification with similar results to those achieved with a medium containing 16% EG-Me2SO.

journal_name

Cryobiology

journal_title

Cryobiology

authors

Cuello C,Sanchez-Osorio J,Almiñana C,Gil MA,Perals ML,Lucas X,Roca J,Vazquez JM,Martinez EA

doi

10.1016/j.cryobiol.2008.02.005

subject

Has Abstract

pub_date

2008-06-01 00:00:00

pages

189-94

issue

3

eissn

0011-2240

issn

1090-2392

pii

S0011-2240(08)00026-6

journal_volume

56

pub_type

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