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You have accessJournal of UrologyInfertility: Physiology, Pathophysiology, Basic Research1 Apr 20112013 INTRODUCTION OF VASA, DAZL, DAZ3, AND BOULE IN THE DIRECT REPROGRAMMING OF GERM CELLS FROM FIBROBLASTS DERIVED FROM ADULT TESTIS TISSUE Hideyuki Kobayashi, Koichi Nagao, Yusuke Oka, Masato Nagata, Fumito Yamabe, keiichiro Takasugi, Shuichi Kamimura, Norie Tanaka, Kuri Suzuki, and Koichi Nakajima Hideyuki KobayashiHideyuki Kobayashi Tokyo, Japan More articles by this author , Koichi NagaoKoichi Nagao Tokyo, Japan More articles by this author , Yusuke OkaYusuke Oka Tokyo, Japan More articles by this author , Masato NagataMasato Nagata Tokyo, Japan More articles by this author , Fumito YamabeFumito Yamabe Tokyo, Japan More articles by this author , keiichiro Takasugikeiichiro Takasugi Tokyo, Japan More articles by this author , Shuichi KamimuraShuichi Kamimura Tokyo, Japan More articles by this author , Norie TanakaNorie Tanaka Tokyo, Japan More articles by this author , Kuri SuzukiKuri Suzuki Tokyo, Japan More articles by this author , and Koichi NakajimaKoichi Nakajima Tokyo, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2241AboutPDF 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. It has been shown that human adult germline stem cells from testicular tissue (haGSCs) have pluripotent properties similar to those of human embryonic stem cells (hESCs). However, recent research has indicated that haGSCs and fibroblasts have a similar gene expression profile and differ in character from hESCs. The pluripotency of haGSCs has thus been questioned. Notably, it was reported that the DAZL, DAZ, and BOULE genes modulate primordial germ cell and haploid gamete formation. Therefore, we examined whether the introduction of VASA, DAZL, DAZ3, and BOULE would permit direct reprogramming of germ cells derived from human testicular fibroblasts. METHODS After obtaining informed consent, testis tissue was collected from infertile men. The tissues were mechanically dissected and dissociated by enzymatic treatment. The cells were then incubated with 7% FBS and DMEM for 14 days. We established fibroblasts from human testicular cultures. VASA, DAZL, DAZ3, and BOULE were delivered to fibroblasts by using a lentiviral vector system. The fibroblasts were then cultured with 7% FBS and DMEM for 4 days, after which they were grown on either a mouse embryonic fibroblast feeder layer, a 0.1% gelatin dish, a HEMA/MMA-coated dish, or an uncoated dish; all dishes were incubated with ES medium and basic fibroblast growth factor. After 20–25 days, cells were collected and their characteristics were investigated by RT-PCR. RESULTS There were no morphological changes in iPS cells generated from testicular fibroblasts with VASA, DAZL, DAZ3, and BOULE; however, RT-PCR confirmed that the cells expressed the germ cell markers VASA, DAZL, and DAZ. CONCLUSIONS We attempted direct reprogramming of germ cells derived from human testicular fibroblasts; however, the introduction of VASA, DAZL, DAZ3, and BOULE did not result in generation of complete germ cells. Nevertheless, we maintain that direct reprogramming of germ cells is possible under suitable conditions. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e805 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hideyuki Kobayashi Tokyo, Japan More articles by this author Koichi Nagao Tokyo, Japan More articles by this author Yusuke Oka Tokyo, Japan More articles by this author Masato Nagata Tokyo, Japan More articles by this author Fumito Yamabe Tokyo, Japan More articles by this author keiichiro Takasugi Tokyo, Japan More articles by this author Shuichi Kamimura Tokyo, Japan More articles by this author Norie Tanaka Tokyo, Japan More articles by this author Kuri Suzuki Tokyo, Japan More articles by this author Koichi Nakajima Tokyo, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...