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Pretreating porcine sperm with lipase enhances developmental competence of embryos produced by intracytoplasmic sperm injection

Published online by Cambridge University Press:  07 October 2015

Yinghui Wei ,
Junhua Fan ,
Lin Li ,
Zhiguo Liu  and
Kui Li
Yinghui Wei
Affiliation:
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China.
Junhua Fan
Affiliation:
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China.
Lin Li
Affiliation:
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China.
Zhiguo Liu*
Affiliation:
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China.
Kui Li
Affiliation:
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China.
*
All correspondence to: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Yuanminyuan West Road No. 2, Haidian District, Beijing 100193, People's Republic of China. Tel: +86 10 62813339. Fax: +86–10–62813339. E-mail: zhiguoliu2010@126.com
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Summary

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.

Keywords

ICSIembryoLipaseTriton X-100
Type
Research Article
Information
Zygote , Volume 24 , Issue 4 , August 2016 , pp. 594 - 602
Copyright
Copyright © Cambridge University Press 2015 

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