Human Reproduction, Vol. 14, No. 2, 458-464,
February 1999
© 1999 European Society of Human Reproduction and Embryology
Development of in-vitro-derived bovine embryos cultured in 5% CO2 in air or in 5% O2, 5% CO2 and 90% N2
1 Department of Reproductive Biotechnology, Louisiana State University–Pennington Biomedical Research Center, Baton Rouge, LA 70808, 2 Department of Animal Science, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA
To evaluate the effects of a three gas mixture of 5% O2, 5% CO2 and 90% N2 (OCN) on preimplantation embryo development, bovine in-vitro fertilization (IVF) oocytes were cultured in a defined medium (mBECM) with various supplements either under 5% CO2 in air or under OCN. When cultured in mBECM alone, embryo development was significantly stimulated in OCN compared to 5% CO2 in air (experiment 1). In the OCN atmosphere, blastocyst formation was further increased after addition of fetal bovine serum (FBS; 10%) or FBS + cumulus granulosa cells (CGC) to mBECM. The ratio of blastocysts to 8-cell embryos, number of hatched blastocysts and embryo diameter were markedly increased, and zona thickness was decreased after FBS addition. However, development up to the morula stage was fully supported by mBECM alone. There was no significant effect of ß-mercaptoethanol (ME; 10 µM) in OCN. In the 5% CO2 atmosphere, embryo development was significantly (P < 0.05) enhanced after addition of FBS + CGC + ME. In experiment 2, in OCN, FBS added at 60 h post-insemination was effective in stimulating blastocyst formation, but changes in medium volume per oocyte from 13.6 to 1.36 µl had only a marginal effect. In conclusion, OCN gas mixture provides a suitable atmosphere for early embryo growth in vitro and mBECM + FBS is the optimal culture medium under this atmosphere.
Key words: bovine/culture atmosphere/defined medium/embryo/IVF
3 Current address: College of Medicine, Pochon CHA University, Infertility Medical Center of CHA General Hospital, 606–5 Yeoksam 1-Dong, Kangnam-Gu, Seoul 135–081, Korea
4 To whom correspondence should be addressed Louisiana State University, Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA 70808-4124, USA
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