title: Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality.
abstract: 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 (∼30 to 50 embryos cultured in 500 μl of CM); Control 5 (Five embryos cultured in 50 μl microdrops of CM), without addition of GF in either of the two control groups; epidermal GF (EGF); IGF-I; and transforming GF-α (TGF-α) (Five embryos were cultured in 50 μl microdrops of CM with 10 ng/ml EGF, 10 ng/ml IGF-I or 10 ng/ml TGF-α, 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 μl 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-α 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 rates and decreasing apoptosis rates of these blastocysts.
authors.affiliation: Centro de Tecnología Animal, Instituto Valenciano de Investigaciones Agrarias, Castellón, Spain.
authors.forename: C J
authors.initials: CJ
authors.lastname: Ahumada
authors.forename: I
authors.initials: I
authors.lastname: Salvador
authors.forename: A
authors.initials: A
authors.lastname: Cebrian-Serrano
authors.forename: R
authors.initials: R
authors.lastname: Lopera
authors.forename: M A
authors.initials: MA
authors.lastname: Silvestre
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002417:Cattle / Q000196:embryology*; D003470:Culture Media / Q000032:analysis*; D003612:Dairying / Q000379:methods; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects*; D004815:Epidermal Growth Factor; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D020287:In Situ Nick-End Labeling / Q000662:veterinary; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D008856:Microscopy, Fluorescence / Q000662:veterinary; D013002:Somatomedins; D015291:Transforming Growth Factors
chemical_list: D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins; D013002:Somatomedins; D004815:Epidermal Growth Factor; D015291:Transforming Growth Factors