Journal
ANIMAL
Volume 7, Issue 3, Pages 455-462Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1751731112001991
Keywords
bovine embryo; reduced groups; in vitro culture; growth factor; apoptosis
Funding
- INIA [RTA2007-0110]
- Conselleria de Educacion of the Generalitat Valenciana [ACOMP/2009/072]
- European Social Fund
- INIA-CCAA
- Ramon y Cajal Subprogramme [RyC-2011-08750]
- Conselleria de Agricultura, Pesca y Alimentacion
- IAMZ-CIHEAM
- FEDER
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When embryos are cultured individually or in small groups, blastocyst yield efficiency and quality are usually reduced. The aim of this work was to investigate the effect of supplementation of the embryo culture medium (CM) with several growth factors (GFs) on embryo development and apoptosis rate when a reduced number of embryos were in vitro cultured. Two experimental studies (ES) were carried out. In ES 1, five treatments were tested to study the effect of GF on embryo development: Control (similar to 30 to 50 embryos cultured in 500 mu l of CM); Control 5 (Five embryos cultured in 50 mu l microdrops of CM), without addition of GF in either of the two control groups; epidermal GF (EGF); IGF-I; and transforming GF-alpha (TGF-alpha) (Five embryos were cultured in 50 mu l microdrops of CM with 10 ng/ml EGF, 10 ng/ml IGF-I or 10 ng/ml TGF-alpha, respectively). In ES 2, following the results obtained in ES 1, four different treatments were tested to study their effect on embryo development and quality (number of cells per blastocyst and apoptotic rate): Control; Control 5; EGF, all three similar to ES 1; EGF + IGF-I group (five embryos cultured in 50 mu l microdrops of CM with 10 ng/ml EGF and 10 ng/ml IGF-I). In both ESs, it was observed that a higher proportion of embryos cultured in larger groups achieved blastocyst stage than embryos cultured in reduced groups (22.6% v. 14.0%, 12.6% and 5.3% for Control v. Control 5, IGF-I, TGF-alpha groups in ES 1, and 24.9% v. 17.1% and 19.0% for Control v. Control 5 and EGF in ES 2, respectively; P < 0.05), with the exception of embryos cultured in medium supplemented with EGF (18.5%) or with EGF + IGF-I (23.5%), in ES 1 and ES 2, respectively. With regard to blastocyst quality, embryos cultured in reduced groups and supplemented with EGF, alone or combined with IGF-I, presented lower apoptosis rates than embryos cultured in reduced groups without GF supplementation (11.6% and 10.5% v. 21.9% for EGF, EGF + IGF-I and Control 5 groups, respectively; P < 0.05). The experimental group did not affect the total number of cells per blastocyst. In conclusion, this study showed that supplementation of the CM with EGF and IGF could partially avoid the deleterious effect of in vitro culture of small groups of bovine embryos, increasing blastocyst and decreasing apoptosis rates of these blastocytes.
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