Abstract: The present invention provides a method of producing a non-human vertebrate that harbours germ cells having a desired gene transferred thereto, comprising injecting the desired gene to the testis of a non-human vertebrate wherein no tight junction exists between Sertoli cells to transfer the desired gene to germ cells, so as to obtain a non-human vertebrate that harbours the germ cells having the desired gene transferred thereto. Using the method of the present invention, even in animal species and lines for which in vitro transduction has been difficult to date, it is possible to obtain an individual harbouring germ cells, particularly spermatogonial stem cells, having a desired gene transferred thereto, at extremely high efficiency.

Abstract: The present invention provides a method of growing spermatogonial stem cells of mammals and the like in vitro, which is characterized in that glial cell-derived neurotrophic factor (GDNF) or an equivalent thereto, and leukemia inhibitory factor (LIF) are contained in a medium (culture broth) for culturing spermatogonial stem cells. According to the method of the present invention, spermatogonial stem cells can be grown in vitro to the extent that enables use thereof for developmental engineering.

Abstract: The present invention provides a method of producing a non-human vertebrate that harbours germ cells having a desired gene transferred thereto, comprising injecting the desired gene to the testis of a non-human vertebrate wherein no tight junction exists between Sertoli cells to transfer the desired gene to germ cells, so as to obtain a non-human vertebrate that harbours the germ cells having the desired gene transferred thereto. Using the method of the present invention, even in animal species and lines for which in vitro transduction has been difficult to date, it is possible to obtain an individual harbouring germ cells, particularly spermatogonial stem cells, having a desired gene transferred thereto, at extremely high efficiency.

Abstract: The present invention provides a method of producing pluripotent stem cells, which comprises culturing testis cells using a medium containing glial cell derived neurotrophic factor (GDNF) or an equivalent thereto to obtain pluripotent stem cells. The medium can further contain leukemia inhibitory factor (LIF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and the like. Using the production method of the present invention, it is possible to produce pluripotent stem cells, which have conventionally been only obtainable from fertilized eggs, embryos and the like, from a postnatal individual. Using the pluripotent stem cells, it is possible to construct diverse tissues having histocompatibility for autotransplantation, and the pluripotent stem cells are useful in medical fields such as regeneration medicine and gene therapy. Also, the pluripotent stem cells are useful in the field of biotechnology because they can be used to prepare transgenic animals, knockout animals and the like.

Abstract: The present invention provides a method of producing offspring originating in a spermatogonial stem cell of a donor animal which comprises generating spermatozoa in the reproductive organ of a male recipient animal with the use of frozen spermatogonial stem cells originating in the donor animal to give a male individual for reproduction, and then producing animal individual originating in the spermatogonial stem cell of the donor using the said individual for reproduction.

Abstract: The present invention provides a method of growing spermatogonial stem cells of mammals and the like in vitro, which is characterized in that glial cell-derived neurotrophic factor (GDNF) or an equivalent thereto, and leukemia inhibitory factor (LIF) are contained in a medium (culture broth) for culturing spermatogonial stem cells. According to the method of the present invention, spermatogonial stem cells can be grown in vitro to the extent that enables use thereof for developmental engineering.