PD39-09 A NOVEL 3-DIMENSIONAL BIOPRINTED MODEL OF HUMAN SPERMATOGENESIS FOR HIGH THROUGHPUT APPLICATIONS

Abstract

You have accessJournal of UrologyInfertility: Basic Research & Pathophysiology (PD39)1 Sep 2021PD39-09 A NOVEL 3-DIMENSIONAL BIOPRINTED MODEL OF HUMAN SPERMATOGENESIS FOR HIGH THROUGHPUT APPLICATIONS Meghan Robinson, Ryan Flannigan, and Luke Witherspoon Meghan RobinsonMeghan Robinson More articles by this author , Ryan FlanniganRyan Flannigan More articles by this author , and Luke WitherspoonLuke Witherspoon More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002049.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Male factor infertility affects 15% of couples, and yet the molecular basis of most cases remains unknown. To address this issue physiologic in vitro models must be developed to investigate the underlying mechanisms of male factor infertility. This study set out to generate a 3-D bioprinted model of spermatogenesis. METHODS: Human testicular biopsies were dissociated and sorted into germ and somatic fractions by differential plating, and expanded. Germ and somatic cells were combined 1:4 and bioprinted into hollow microfibers using an Aspect Biosystems RX1 bioprinter with a coaxial printhead. Bioprinted fibers were placed in media with Epidermal Growth Factor (EGF), Leukemia Inhibitory Factor (LIF), Bone Morphogenic Protein 4 (BMP4), Stem Cell Factor (SCF), Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), metribolone, and retinoic acid, and incubated at 5% CO2 and 34°C for 12 days. Live-dead staining was performed with SYBR14 and ethidium homodimer dyes. Gene expression was assessed by real time quantitative polymerase reaction. Organization was analyzed using hematoxylin and eosin staining. RESULTS: Human primary testicular cells proved to be expandable in culture, and highly viable after bioprinting (Figure 1B). Sertoli cell cords formed within the bioprinted fibers and the organoids, illustrating an intrinsic ability to self-organize (Figure 1D-E). Germ cells in the bioprinted fibers upregulated gene expression associated with spermatogonial stem cell quiescence, proliferation, meiosis, and spermiogenesis (Figure 1F-G). Overall, their ability to self-organize and support spermatogenic activity was on par with organoids. CONCLUSIONS: This study provides a proof-of-principle 3-D bioprinted models of spermatogenesis. We show that bioprinted testicular cells retain their phenotypes and functionality, opening the door to high throughput applications. Source of Funding: VCHRI, CUASF, CIHR, UBC. © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e671-e671 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Meghan Robinson More articles by this author Ryan Flannigan More articles by this author Luke Witherspoon More articles by this author Expand All Advertisement Loading ...