A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes.
The general method of porcine in vitro fertilization (IVF), involving the co-culture of both gametes in a medium drop, is thought to be the main reason for the high incidence of polyspermy. The aim of this study was to reduce the polyspermic fertilization of porcine embryos during IVF by the modified swim-up method, based on general sperm swim-up technique. Within this design, a 70 microm pore sized cell strainer was used to separate the sperm pellet placed at the bottom of a tube from the mature oocytes placed within the upper region. The separation of gametes using this permeable barrier was to ensure that only motile sperm gained access to the oocytes. It was found that the rate of polyspermy was significantly lowered for the sperm preparations from three boar breeds in modified swim-up method when compared with that of the general microdrop method (p<0.05). However, the penetration rates were found to be similar in both methods for two boar breeds. The average occurrence of blastocysts with more total cell number was higher in the modified swim-up method, while no significant difference in blastocyst rates between the two IVF methods was observed. The frequency of normal diploid embryos was also significantly higher in the modified swim-up method and polyploidy was more frequently observed in microdrop method (p<0.05). Our results demonstrated that the modified swim-up IVF method could reduce polyspermic penetration, and consequently produce better quality and karyotypically normal embryos in porcine IVF.
Park
Chi-Hun
CH
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-921, Republic of Korea.
Lee
Sang-Goo
SG
Choi
Don-Ho
DH
Lee
Chang-Kyu
CK
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary; D004542:Ejaculation; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005307:Fertilization in Vitro / Q000379:methods / Q000592:standards / Q000662:veterinary*; D007621:Karyotyping; D008297:Male; D009865:Oocytes / Q000502:physiology*; D011003:Ploidies; D011247:Pregnancy; D012661:Semen / Q000502:physiology; D013079:Sperm Maturation / Q000502:physiology; D013081:Sperm Motility / Q000502:physiology*; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000166:cytology; D013552:Swine; D015044:Zona Pellucida / Q000502:physiology / Q000648:ultrastructure