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Human Reproduction, Vol. 14, No. 11, 2838-2843, November 1999
© 1999 European Society of Human Reproduction and Embryology

Simple, efficient and successful vitrification of bovine blastocysts using electron microscope grids

Se-Pill Park1,5, Eun Young Kim1, Deok Im Kim2, Noh Hyung Park2, Yu Seok Won2, San Hyun Yoon3, Kil Saeng Chung4 and Jin Ho Lim3

1 Maria Infertility Medical Institute, 103–11 Sinseol-dong Dongdaemun-Gu, Seoul 130–110, 2 Hanwoo Improvement Center, NLCF, Seosan, Chungnam 356–830, 3 Maria Infertility Clinic, Sinseol-dong Dongdaemun-Gu, Seoul 130–110, 4 Department of Animal Sciences, Kon-Kuk University, Mojin-dong, Kwangjin-gu, Seoul 143–701, Korea

This study demonstrates that higher survival of vitrified–thawed bovine blastocysts can be obtained using electron microscope (EM) grids as embryo containers at freezing, rather than plastic straws. In-vitro produced day 7 bovine blastocysts after in-vitro fertilization (IVF) were vitrified on grids or in straws with EFS40 freezing solution and their survival after thawing was compared. Embryo survival was assessed as re-expanded and hatched rates at 24 and 48 h after thawing respectively. When the effects of exposure to vitrification solution and chilling injury from the freezing procedure were examined, embryo survival in the exposure group (24 h: 100, 48 h: 73.3%) was not different compared with that in the control group (100, 84.4%). After vitrification, the hatched rate of the EM grid group 48 h after thawing (67.8%) was significantly higher than that of the straw group (53.3%) (P < 0.05). Fast developing embryos (expanded blastocyst and early hatching blastocyst stage) showed better resistance to freezing than delayed ones (early blastocyst stage), irrespective of embryo containers (early: 24 h, 57.1 and 48 h, 24.4%; expanded: 84.7 and 60.6%; early hatching: 91.7 and 80.0%) (P < 0.001). When using expanded and early hatching blastocysts, embryo survival rates in the vitrification-EM grid group (67.8, 95.0% respectively) were significantly higher than that of the vitrification-straw group (53.0, 65.0%) at 48 h.

Key words: bovine blastocysts/in-vitro survival/EM grid/plastic straw/vitrification

5 To whom correspondence should be addressed


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