Abstract
This study aimed to compare the quality and the penetration ability of frozen–thawed spermatozoa from three microminipigs and Large White boars and to evaluate the effects of caffeine and heparin as well as the sperm–oocyte co-incubation length on the fertilization and embryonic development in vitro. Results showed that the fertilization rates of spermatozoa from three microminipig boars were significantly lower than those of a Large White boar. In the post-thaw spermatozoa from one of three microminipig boars, the sperm quality, penetration ability, and the oocyte development after in vitro fertilization were significantly lower than those of the spermatozoa from other boars. The caffeine supplementation in the fertilization media increased the rates of fertilization and blastocyst formation for the microminipig spermatozoa with low sperm quality. In addition to caffeine supplementation, the rates of fertilization and blastocyst formation after using microminipig spermatozoa were significantly higher with a 10 h sperm–oocyte co-incubation than with 3 h of co-incubation length. Our results indicate that the differences between the males and the breed influence the quality and fertility of frozen–thawed boar spermatozoa. In conclusion, the presence of caffeine in the in vitro fertilization (IVF) medium and adequate length of sperm–oocyte co-incubation may have beneficial effects for improving IVF results when using microminipig spermatozoa with low quality.
Highlights
Microminipigs have a tiny body size (7 to 8 kg for a 6-monthold mature pig), which provides many advantages for laboratory use, including ease of handling, small rearing spaces, and low doses of test substances (Kaneko et al, 2011)
To improve the poor penetration ability of microminipig spermatozoa, we evaluated the effects of caffeine and heparin supplementation during in vitro fertilization (IVF) and sperm–oocyte co-incubation length on the fertilization and development of porcine oocytes
Semen was collected from three microminipig boars (1.5–2 years old) with unknown results of sperm parameter, fertilization, and artificial insemination (AI) and from a Large White boar (3 years old) with known in vitro fertilization results but unknown sperm parameter and AI results, using the “glovedhand” technique
Summary
Microminipigs have a tiny body size (7 to 8 kg for a 6-monthold mature pig), which provides many advantages for laboratory use, including ease of handling, small rearing spaces, and low doses of test substances (Kaneko et al, 2011). A cross between microminipigs and the large western pig is expected to result in a smaller pig than the western pig This method is useful, for example, when miniaturizing genetically modified animals produced from large western pigs. The natural mating of microminipigs with the large western pig is difficult due to differences in size. The use of in vitro fertilization (IVF) is one of the useful tools for the production of a crossbreed using frozen–thawed spermatozoa of microminipigs as an alternative to natural mating. The freezing and thawing processes result in compromised sperm function and IVF success. It has been suggested that various circumstances are associated with the low fertility of frozen– thawed boar spermatozoa, which include damage to the spermatozoa during the freeze–thaw process (Yeste, 2016).
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