Optimization of the in vitro fertilization protocol for frozen epididymal sperm with low fertilization ability in Ban—A native Vietnamese pigs |
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| Authors: | Nguyen Viet Linh Tamas Somfai Thi Hiep Nguyen Nguyen Thi Nhung Nguyen Thi Hong Nguyen Tien Dat Nguyen Hoang Thinh Nguyen Khanh Van Dong Van Quyen Hoang Ha Chu Nguyen Thanh Son Kazuhiro Kikuchi |
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| Affiliation: | 1. Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi, Vietnam;2. Graduate University of Science and Technology, Vietnam Academy of Science and Technology, Hanoi, Vietnam;3. Division of Animal Breeding and Reproduction Research, Institute of Livestock and Grassland Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan;4. Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan;5. The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi, Yamaguchi, Japan;6. Faculty of Animal Science, Vietnam National University of Agriculture, Hanoi, Vietnam;7. National Institute of Animal Sciences, Hanoi, Vietnam |
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| Abstract: | The aim of the present study was to improve the penetration during in vitro fertilization (IVF) of a frozen lot of epididymal sperm with a notoriously low fertilization ability of a Ban boar which is a native Vietnamese breed by optimizing different parameters of the IVF system. In Experiment 1, we determined that Pig‐fertilization medium was superior medium to Tyrode's albumin lactate pyruvate‐polyvinyl alcohol medium for IVF and defined the optimum the sperm concentration (1 × 106 sperm/ml). In Experiment 2, we clarified that partial removal of cumulus cells from cumulus‐oocyte complexes by hyaluronidase treatment before IVF enhances sperm penetration, whereas complete cumulus removal reduces penetration. Finally, in Experiment 3 the elevation of concentration of caffeine in Pig‐fertilization medium from 2 to 5 mmol/L and the prolongation of the co‐culture of gametes from 3 to 5 hr significantly increased the total penetration rate from 15.2% to over 50%. In conclusion, the combination of partial oocyte denudation, an elevated caffeine concentration in Pig‐fertilization medium and an extended interval of IVF with using an optimized sperm concentration was a potent way to improve the fertilization results for a frozen epididymal Ban sperm lot with low fertility. |
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| Keywords: | Ban breed epididymal sperm freezing in vitro fertilization Vietnamese native boar |
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