Earticle

초록

영어

Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: In vitro -matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 μL microdroplets (10 gametes/drop) or 500 μL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Oocyte collection and in vitro maturation
Sperm preparation
In vitro fertilization and in vitro culture
Assessment of fertilization parameters
Embryo evaluation and total cell count
Experimental design
Statistical analysis
RESULTS
Effects of sperm-free culture time on polyspermy in porcine embryos
Effect of sperm-oocyte co-culture time on development and quality of embryos
Effect of Percoll treatment of sperm on penetration and blastocyst formation in embry
Effect of volume of culture medium on survivability and development of embryos
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• Song-Hee Lee [ Department of Animal Science, Chungbuk National University, Cheongju 28644, Korea ]
• Xiang-Shun Cui [ Department of Animal Science, Chungbuk National University, Cheongju 28644, Korea ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2025

• 십진분류

  KDC 527 DDC 636