title: Adenovirus-mediated gene delivery into mouse spermatogonial stem cells.
abstract: Spermatogonial stem cells represent a self-renewing population of spermatogonia, and continuous division of these cells supports spermatogenesis throughout the life of adult male animals. Previous attempts to introduce adenovirus vectors into spermatogenic cells, including spermatogonial stem cells, have failed to yield evidence of infection, suggesting that male germ cells may be resistant to adenovirus infection. In this study we show the feasibility of transducing spermatogonial stem cells by adenovirus vectors. When testis cells from ROSA26 Cre reporter mice were incubated in vitro with a Cre-expressing adenovirus vector, Cre-mediated recombination occurred at an efficiency of 49-76%, and the infected spermatogonial stem cells could reinitiate spermatogenesis after transplantation into seminiferous tubules of infertile recipient testes. No evidence of germ-line integration of adenovirus vector could be found in offspring from infected stem cells that underwent Cre-mediated recombination, which suggests that the adenovirus vector infected the cells but did not stably integrate into the germ line. Nevertheless, these results suggest that adenovirus may inadvertently integrate into the patient's germ line and indicate that there is no barrier to adenovirus infection in spermatogonial stem cells.
authors.affiliation: Department of Molecular Genetics, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Takehashi
authors.forename: Mito
authors.initials: M
authors.lastname: Kanatsu-Shinohara
authors.forename: Kimiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.forename: Hiromi
authors.initials: H
authors.lastname: Miki
authors.forename: Shinya
authors.initials: S
authors.lastname: Toyokuni
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
mesh_terms: D000256:Adenoviridae / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D005260:Female; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D006801:Humans; D007314:Insemination; D019426:Integrases / Q000378:metabolism; D008297:Male; D051379:Mice; D011506:Proteins / Q000235:genetics; D022661:RNA, Untranslated; D011995:Recombination, Genetic; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D016662:Virus Integration; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D011506:Proteins; D022661:RNA, Untranslated; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase