PD39-06 UTILIZATION OF IMMUNOHISTOCHEMISTRY AS A MEANS TO IDENTIFY SPERMATOGONIAL STEM CELLS IN TESE NEGATIVE KLINEFELTER SYNDROME PATIENTS: BUILDING BLOCKS FOR IN VITRO SPERMATOGENESIS? (2021)

You have accessJournal of UrologyInfertility: Basic Research & Pathophysiology (PD39)1 Sep 2021PD39-06 UTILIZATION OF IMMUNOHISTOCHEMISTRY AS A MEANS TO IDENTIFY SPERMATOGONIAL STEM CELLS IN TESE NEGATIVE KLINEFELTER SYNDROME PATIENTS: BUILDING BLOCKS FOR IN VITRO SPERMATOGENESIS? Nicholas Deebel, Haleh Soltangoraee, Karl Reynolds, Kimberly Stogner-Underwood, Mohamad Reza Sadeghi, Anthony Atala, and Hooman Sadri-Ardekani Nicholas DeebelNicholas Deebel More articles by this author , Haleh SoltangoraeeHaleh Soltangoraee More articles by this author , Karl ReynoldsKarl Reynolds More articles by this author , Kimberly Stogner-UnderwoodKimberly Stogner-Underwood More articles by this author , Mohamad Reza SadeghiMohamad Reza Sadeghi More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and Hooman Sadri-ArdekaniHooman Sadri-Ardekani More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002049.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Klinefelter Syndrome (KS) is defined by chromosomal aneuploidy (typically 47, XXY). KS is associated with progressive testicular fibrosis, loss of spermatogonial stem cells (SSC), and impaired fertility. More than 50% of KS patients will be negative for spermatozoa on testicular sperm extraction (TESE). Therefore, therapies utilizing stem cell therapy i.e. SSCs have been explored. In order to assess the practicality of these therapies, developing a clinically validated method to detect SSCs and defining their prevalence in KS is needed. METHODS: KS patients (n=79) underwent bilateral TESE as an initial effort to recover spermatozoa for IVF/ICSI. Control samples (n=12) were obtained from brain dead, non-Klinefelter Syndrome patients. The institution’s IRB approved this study. A dedicated genitourinary pathologist examined the H&E stained tissue for the presence of spermatogonia and spermatozoa. An extra slide from each patient was stained in an automated, clinically validated system with PGP9.5 (UCHL1) antibody and MAGE-A4 antibody, specific markers for undifferentiated spermatogonia in seminiferous tubules. The stained slides were analyzed with virtual microscopy. In addition, clinical data such as age, karyotype, height, weight, average testicle size, and hormonal panel (LH, FSH, Testosterone) were collected. RESULTS: PGP9.5 staining showed that 74.7% (n= 59) of the KS patients were positive for undifferentiated spermatogonia compared to 100% (n=12) in the control patients (p <0.05). MAGE-A4 staining showed similar results. H&E with microscopic analysis showed only 8.9% (n=7) of the patients were positive for spermatogonia (p <0.0001). The mean number of positive tubules per square millimeter for the control and KS patients were 11.9±1.88 and 1.3±0.14 respectively (p <0.0001). Additionally, PGP9.5 and MAGE-A4 staining showed that more than 70% of the spermatozoa negative patients (on TESE), still had spermatogonia. Multivariate analysis with linear regression showed no significant correlation between clinical variables and the number of PGP9.5 positive tubules found on biopsy. CONCLUSIONS: Conclusions: Either PGP9.5 or MAGE-A4 immunostaining are an effective way of identifying undifferentiated spermatogonia in KS patients. Many patients remain positive for SSCs which could be harvested and used for potential infertility therapy such as SSC in vitro propagation and differentiation followed by In vitro fertilization. Source of Funding: NA © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e670-e670 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nicholas Deebel More articles by this author Haleh Soltangoraee More articles by this author Karl Reynolds More articles by this author Kimberly Stogner-Underwood More articles by this author Mohamad Reza Sadeghi More articles by this author Anthony Atala More articles by this author Hooman Sadri-Ardekani More articles by this author Expand All Advertisement PDF downloadLoading ...