MP60-15 TRANSCRIPTOME ANALYSIS TO IDENTIFY HUMAN SPERMATOGONIAL CELLS FROM SERTOLI CELL-ONLY TESTES

Abstract

You have accessJournal of UrologyInfertility: Basic Research & Pathophysiology1 Apr 2018MP60-15 TRANSCRIPTOME ANALYSIS TO IDENTIFY HUMAN SPERMATOGONIAL CELLS FROM SERTOLI CELL-ONLY TESTES Koji Shiraishi and Hideyasu Matsuyama Koji ShiraishiKoji Shiraishi More articles by this author and Hideyasu MatsuyamaHideyasu Matsuyama More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1907AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Non-obstructive azoospermia (NOA), particularly Sertoli cell-only (SCO), is the most severe form of male factor infertility and is classically thought to lack germ cells and spermatogenic stem cells (SSCs). However, evidence from microsurgical testicular sperm extraction (micro-TESE) on men with SCO has revealed the presence of focal areas of full spermatogenesis, suggesting the possible presence of SSCs in some patients with SCO lacking mature sperm. To determine whether gene products from SSCs could be detected in SCO, transcriptome analysis using next-generation sequencing (NGS) was performed using human biopsied testicular samples. METHODS Under IRB approval, testicular samples were obtained from 20 men (mean age: 35.7 years) with SCO in which no germ cells were found during laboratory examination and from five men with obstructive azoospermia (mean age: 38.5 years) with normal spermatogenesis. Men with sex chromosome or AZF abnormality were excluded. Transcriptomes on the Illumina platform were expressed as FPKM, and Cuffdiff analysis was performed to compare individual differences among testicular tissues from obstructive azoospermia (OA). Expression of mRNA and localization of targeted genes were confirmed by RT-PCR and immunohistochemical analyses, respectively. RESULTS From the 23,003 genes screened with using NGS with a significant signal after background correction and normalization, a total of 5,666 genes were analyzed. As representative markers of immature germ cells, UTF1, OCT4, DDX4, MMP1, GFRα, GPX1, SCP3, CD14, KLF9, GFRA1 and KIT mRNA were detected in SCO samples by transcriptome analysis. RT-PCR showed significant mRNA expression of UTF1, OCT4, DDX4, SCP3 and KIT from SCO samples. Immunohistochemistry showed the localization of DDX4 and/or KIT-positive cells in the seminiferous tubules and basement membrane in eight (40%) of the SCO samples. All the gene products evaluated in SCO samples were present in testes with obstructive azoospermia. There was no significant difference in the patients' background and hormonal data between the men with and without DDX4 and/or KIT-positive cells. CONCLUSIONS Germ cells are present in 40% of SCO samples as immature forms, namely SSCs. These observations help our understanding of the pathophysiology of spermatogenic failure where germ cell development stops during early stages of spermatogenesis in some men with SCO. The azoospermia seen in men with SCO appears to be, at least in part, secondary to a defect in proliferation and differentiation of SSCs. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e795 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Koji Shiraishi More articles by this author Hideyasu Matsuyama More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...