J Vet Sci.  2003 Apr;4(1):73-78.

Effects of Protein Source and Energy Substrates on the In Vitro Development of Bovine Embryos in a Two-step Culture System

Affiliations
  • 1Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. firstlee@snu.ac.kr

Abstract

In this study, we examined the effects of a two-step culture system, which involves the use of different culture media for early cleavage and later stage embryos, on the in vitro development of bovine embryos. We also investigated the effect of glucose, phosphate and citrate on the in vitro early developmental period of bovine embryos in a two-step culture system. Moreover, the supplementation of different protein sources (BSA-V, BSA-FAF and FBS) during IVC did not affect the frequency of blastocyst development. Using two-step culture, embryos were cultured in protein-free media for an initial 5 days. This was then followed by the same culture media or an FBS supplemented media. The developmental rates of blastocysts in the FBS containing group were significantly higher than in the replaced with no serum containing group. Embryos cultured in mSOF supplemented with 1.5 mM glucose plus 1.2 mM phosphate were significantly inhibited. The inhibition of developmental competence by glucose plus phosphate was consistent with the existence of 0.5 mM sodium citrate. This study indicates that a two-step culture system, which applies different conditions for early cleavage embryos, i.e., serum-free media, vs. later stage embryos, with serum containing media, may be effective for in vitro production systems. In addition, the developmental competence of bovine embryos was depressed in the presence of glucose plus phosphate as compared to either alone or the absence of both. Therefore, the avoidance of this negative effect should allow more optimal conditions to be developed for in vitro production.

Keyword

embryo; in vitro development; two-step culture system; glucose; phosphate; citrate

MeSH Terms

Animals
Blastocyst/drug effects/metabolism
Cattle/*embryology
Citric Acid/pharmacology
Culture Media/*chemistry
Culture Techniques/*methods
Ectogenesis/drug effects
Embryo/*drug effects/embryology/metabolism
Embryonic and Fetal Development/drug effects
*Energy Metabolism
Female
Fertilization in Vitro
Glucose/pharmacology
Male
Phosphates/pharmacology
Proteins/*pharmacology
Zygote/drug effects/metabolism
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