Abstract
Evaporative drying (ED) is an alternative technique for long-term preservation of mammalian sperm, which does not require liquid nitrogen or freeze-drying equipment, but offers advantages for storage and shipping at ambient temperature and low cost. However, the development of zygotes generated from these sperms was poor. Here, we demonstrated that the supplementation of tauroursodeoxycholic acid (TUDCA), an endogenous bile acid, during embryo culture improved the developmental competency of embryos derived from in vitro matured pig oocytes injected intracytoplasmically with boar ED spermatozoa by reducing the production of reactive oxygen species, the DNA degradation and fragmentation, and the expression of apoptosis-related gene Bax and Bak, and by increasing the transcription of anti-apoptosis gene Bcl-XL and Bcl-2. Furthermore, TUDCA treatment promoted the blastocyst quality manifested by the total cell numbers and the ratio of inner cell mass. Taken together, our data suggest that evaporative drying would be a potentially useful method for the routine preservation of boar sperm in combination with further optimization of subsequently embryo culture conditions.
Highlights
Long-term preservation of mammalian sperm has widespread applications in medicine and agriculture, including facilitating treatment of human infertility and fertility preservation for male cancer patients, artificial insemination and genetic improvement of livestock, conservation of endangered species and maintenance of genetic diversity
To evaluate the feasibility that storage of boar sperm through evaporative-drying technique, we examined the development of embryos derived by Intracytoplasmic sperm injection (ICSI) of evaporatively dried spermatozoa following porcine cumulus-oocyte complexes (COCs) maturation in vitro
In consistent with the rate of expanded and hatched blastocyst, the quality of embryos manifested by the total cell numbers and the ratio of inner cell mass was significantly higher in 200 μM tauroursodeoxycholic acid (TUDCA) treatment group than that in without TUDCA treatment group and cryopreserved spermatozoa group (N = 3, p < 0.05) (Table 3)
Summary
Long-term preservation of mammalian sperm has widespread applications in medicine and agriculture, including facilitating treatment of human infertility and fertility preservation for male cancer patients, artificial insemination and genetic improvement of livestock, conservation of endangered species and maintenance of genetic diversity. While cryopreservation of mammalian sperm in liquid nitrogen (LN2) has been successfully employed, the ability to preserve sperm at much higher temperatures (ideally at ambient temperature) is preferred for a number of reasons. It would dramatically reduce the storage expenses, facilitate sample transport while reducing associated costs, and prevent cross contamination between samples stored in LN21–4. It is tempting to speculate that addition of antioxidant TUDCA during embryo culture to reduce the levels of ROS in embryos derived from evaporative drying spermatozoa can improve the poor outcome following ICSI. Our results demonstrate that TUDCA treatment increased the development and the quality of embryos from spermatozoa evaporatively dried in swine
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