title: Pretreating porcine sperm with lipase enhances developmental competence of embryos produced by intracytoplasmic sperm injection.
abstract: Intracytoplasmic sperm injection (ICSI) has been widely applied in humans, mice, and some domestic animals to cure human infertility, or produce genetically superior or genetically engineered animals. However, the production efficiency of ICSI in pigs remains quite low. In this study, we developed a new sperm pretreatment method to improve production efficiency of ICSI in pigs. Experiment 1 revealed that pretreating porcine sperm with 2.5 mg/ml lipase before ICSI operation, not only can reduce the adhesion between sperm and the injection pipette without adding polyvinylpyrrolidone (PVP) in the operating medium, but also significantly improve male pronuclei (MPN) formation rate (55.56% vs. 40.00% (0 mg/ml), 42.59% (5.0 mg/ml), 40.00% (10.0 mg/ml), P < 0.05) and enhance developmental competence of ICSI embryos (26.03% vs. 10.87% (0 mg/ml), 10.00% (5.0 mg/ml), 10.13% (10.0 mg/ml), P < 0.05). Experiment 2 showed that this method has a higher MPN formation rate (50.47% vs. 30.78%, P < 0.05) and blastocyst rate (18.81% vs. 7.41%, P < 0.05) than the PVP method, and was better than the Triton X-100 treatment method (50.47% vs. 46.23%, 18.81% vs. 12.75%). Therefore, pretreating porcine sperm with 2.5 mg/ml lipase before ICSI operation is highly recommended, instead of adding PVP in the operating medium.
authors.affiliation: Institute of Animal Science,Chinese Academy of Agricultural Sciences,Yuanminyuan West Road No. 2,Haidian District,Beijing 100193,People's Republic of China.
authors.forename: Yinghui
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Institute of Animal Science,Chinese Academy of Agricultural Sciences,Yuanminyuan West Road No. 2,Haidian District,Beijing 100193,People's Republic of China.
authors.forename: Junhua
authors.initials: J
authors.lastname: Fan
authors.affiliation: Institute of Animal Science,Chinese Academy of Agricultural Sciences,Yuanminyuan West Road No. 2,Haidian District,Beijing 100193,People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: Institute of Animal Science,Chinese Academy of Agricultural Sciences,Yuanminyuan West Road No. 2,Haidian District,Beijing 100193,People's Republic of China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Institute of Animal Science,Chinese Academy of Agricultural Sciences,Yuanminyuan West Road No. 2,Haidian District,Beijing 100193,People's Republic of China.
authors.forename: Kui
authors.initials: K
authors.lastname: Li
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000502:physiology; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology; D005260:Female; D059471:In Vitro Oocyte Maturation Techniques; D008049:Lipase / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology; D013552:Swine
chemical_list: D008049:Lipase
keywords: ICSI; Lipase; Triton X-100; embryo