ARS | Publication request: The Effect of Culture Methods and Serum Supplementation on Developmental Competence of Bovine Embryos Cultured In Vitro

Submitted to: Journal of Animal and Veterinary Advances
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 22, 2009
Publication Date: January 1, 2009
Citation: Jung, S.Y., Willard, S.T. 2009. The Effect of Culture Methods and Serum Supplementation on Developmental Competence of Bovine Embryos Cultured In Vitro. Journal of Animal and Veterinary Advances. 8(1):67-74.

Interpretive Summary: Bovine embryo culture in the laboratory is sometimes difficult, and efforts are being made to try and evaluate alternate environments to improve embryo fertilization rates and the culture environment for improved embryo production. The objective of this study was to compare the developmental competence of bovine in vitro fertilized embryos in three different culture methods: the microdrop method, the tube method and with or without oil overlay and fetal bovine serum as a supplement to the medium. Overall, the microdrop method with oil overlay was the optimum culture method among the culture methods tested, and serum supplementation did not significantly affect the blastocyst rate. These findings may be used to establish the optimal culture methods for embryos in the laboratory, with potential implications to embryo holding, culture and transfer as related to the production environment.

Technical Abstract: The objective of this study was to compare the developmental competence of bovine in vitro fertilized embryos in three different culture methods; microdrop method (50 µl of medium under mineral oil in petri dishes) compared to tube methods (1 ml of medium in tubes) with or without oil overlay, and to examine the influence of fetal bovine serum (FBS) in culture methods. They were randomly allocated to one of following culture conditions: (Group 1) microdrop method supplemented with 10% (v:v) FBS at day 5 post-insemination (pi); (Group 2) microdrop method without FBS; (Group 3) tube method with oil overlay and FBS added at day 5 pi; (Group 4) tube method with oil overlay; (Group 5) tube method without oil overlay but FBS added at day 5 pi; (Group 6) tube method without oil overlay and FBS. There were no differences (P >0.05) in cleavage rates among the culture methods (78.3%, 75.6%, and 74.9% for microdrop, tube with oil overlay, and tube method without overlay, respectively). However, regardless of serum addition, blastocyst rates in the microdrop method (30.5%) were significantly higher (P <0.05) than those in the tube method without oil overlay (9.8%). There was no difference (P >0.05) between with or without serum in blastocyst rate regardless culture methods. Numerically, the highest blastocyst rate was observed in the microdrop method with the FBS supplementation (Group 1; 36.6%). Overall, the microdrop method was the optimum culture method among the culture methods; however, serum supplementation did not significantly affect the blatocyst rate.