1877 use of ips technology to generate pluripotent stem cells derived from adult testis tissue from infertile men

You have accessJournal of UrologyInfertility: Physiology, Pathophysiology, Basic Research1 Apr 20101877 USE OF IPS TECHNOLOGY TO GENERATE PLURIPOTENT STEM CELLS DERIVED FROM ADULT TESTIS TISSUE FROM INFERTILE MEN Hideyuki Kobayashi, Yusuke Oka, Koichi Nagao, Takao Ishiwatari, Shigeki Kanbe, Masato Nagata, Fumito Yamabe, Keiichiro Takasugi, Taisuke Suyama, Genichi Ishii, Kuri Suzuki, Norie Tanaka, Hiroshi Hara, and Nobuhisa Ishii Hideyuki KobayashiHideyuki Kobayashi More articles by this author , Yusuke OkaYusuke Oka More articles by this author , Koichi NagaoKoichi Nagao More articles by this author , Takao IshiwatariTakao Ishiwatari More articles by this author , Shigeki KanbeShigeki Kanbe More articles by this author , Masato NagataMasato Nagata More articles by this author , Fumito YamabeFumito Yamabe More articles by this author , Keiichiro TakasugiKeiichiro Takasugi More articles by this author , Taisuke SuyamaTaisuke Suyama More articles by this author , Genichi IshiiGenichi Ishii More articles by this author , Kuri SuzukiKuri Suzuki More articles by this author , Norie TanakaNorie Tanaka More articles by this author , Hiroshi HaraHiroshi Hara More articles by this author , and Nobuhisa IshiiNobuhisa Ishii More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1830AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The forced expression of 4 transcription factors—Oct4, Sox2, Klf4, and c-Myc—permits reprogramming of fibroblasts into pluripotent stem cells. These reprogrammed cells are referred to as induced pluripotent stem (iPS) cells. Recent research has shown that pluripotent stem cells can be generated from adult human testis, and it has been suggested that culture conditions are particularly important for spermatogonial stem cells (SSCs) and precursors, after reprogramming of pluripotent stem cells. However, complete reprogramming of cells derived from testis tissue is difficult even under ideal culture conditions. In addition, these stem cells require SSCs or precursors in the testis to produce normal spermatogenesis. We examined whether iPS technology would permit generation of pluripotent stem cells from adult testis tissue collected from infertile men. METHODS After obtaining informed consent, testis tissue was collected from patients with suspected male infertility (patients with oligospermia and 1 patient with Klinefelter syndrome). Tissues were mechanically dissected and dissociated by enzymatic treatment. The cells were then incubated with 7% FBS and DMEM for 4 to 5 days. Oct4, Sox2, Klf4, and c-Myc were delivered to testis cells by using a lentiviral vector system. Testis cells were then cultured with 7% FBS and DMEM for 4 days. Next, testis cells were grown on a mouse embryonic fibroblast feeder layer and incubated with ES medium and basic fibroblast growth factor. After 16 days, a small number of colonies were visible by microscopy. After 20-25 days, ES-like colonies were collected and their characteristics were investigated. RESULTS We were able to collect ES-like cells derived from infertile men. Short tandem repeat analysis confirmed that these colonies were derived from testis tissue. ES-like cells from the patient with Klinefelter syndrome were examine by G-band analysis: 47,XXY was present. In patients without Klinefelter syndrome, testis cells expressed OCT4, NANOG, STELLA, VASA, and DAZL. The colonies expressed OCT4, NANOG, and STELLA, but did not express the germ cell markers VASA and DAZL. We used embryoid body formation and teratoma formation to assess pluripotency both in vitro and in vivo. CONCLUSIONS Using iPS technology, pluripotent stem cells can be generated from adult testis tissue from infertile men. Analysis of iPS cells from such patients may help to determine the mechanisms underlying male infertility. Tokyo, Japan© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e729-e730 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hideyuki Kobayashi More articles by this author Yusuke Oka More articles by this author Koichi Nagao More articles by this author Takao Ishiwatari More articles by this author Shigeki Kanbe More articles by this author Masato Nagata More articles by this author Fumito Yamabe More articles by this author Keiichiro Takasugi More articles by this author Taisuke Suyama More articles by this author Genichi Ishii More articles by this author Kuri Suzuki More articles by this author Norie Tanaka More articles by this author Hiroshi Hara More articles by this author Nobuhisa Ishii More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...