Hiroshima University

Renia Medical Group

Masayuki Shimada

Tetsuji Okazaki

Takashi Umehara

Masaaki Goto

Moeka Kugumiya

Yoko Fujita

Provided are a method for separating mammalian sperm, by which even a large amount of sperm can be separated in a short time and decrease in motility of the separated sperm can be prevented; and an artificial insemination method, and an in vitro fertilization method. The method for separating mammalian sperm includes incubating the sperm in a medium containing a TLR7 ligand, and separating them into sperm floating in the upper layer and sperm sinking in the lower layer of the medium. The artificial insemination and in vitro fertilization methods include artificial insemination or in vitro fertilization for a non-human mammal using sperm of the non-human mammal separated by the method for separating mammalian sperm.

Method for separating mammalian sperm, artificial insemination method, and in vitro fertilization method

A sperm diluent of the present invention contains a chelating agent such as EDTA and/or, EGTA, which forms a complex with a calcium ion, in a base diluent. Further, the sperm diluent contains an immunosuppressive factor such as a steroid hormone and/or, a cytokine, which suppresses migration of leukocytes. By diluting frozen sperm with this sperm diluent followed by performing artificial insemination, death of the sperm before fertilization and phagocytosis of the sperm and embryos by leukocytes in the uterus can be suppressed, allowing enhancement of the conception rate and the implantation rate.

Sperm diluent solution and method for artificial insemination using same

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Higashihiroshima, Japan

Masayuki Shimada