Xeno-free peptide-functionalized bioinks improve ex vivo culture of human testicular tissues

You have accessJournal of UrologyCME1 Apr 2023MP01-03 XENO-FREE PEPTIDE-FUNCTIONALIZED BIOINKS IMPROVE EX VIVO CULTURE OF HUMAN TESTICULAR TISSUES Meghan Robinson and Ryan Flannigan Meghan RobinsonMeghan Robinson More articles by this author and Ryan FlanniganRyan Flannigan More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003212.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The development of human in vitro spermatogenesis (IVS) is a promising regenerative platform for the treatment of male factor infertility. IVS has been demonstrated in rodent models, but despite decades of research remains a challenge in the human sphere. A promising strategy may be to culture fragments of testis tissues in hydrogels. Hydrogels can be functionalized to sequester matrix and signaling peptides in a local manner mimicking tissue niche function. In this study we investigated the use of xeno-free hydrogels functionalized with biomimetic peptide motifs for their ability to improve ex vivo culture of human testicular tissues. METHODS: Human testicular tissues were obtained from hormone-treated gender affirmation patients following informed consent, gently teased apart, cut into small fragments and mixed into bioinks (VitroInk) containing either YIGSR (laminin), COL (collagen), or MMPs (matrix metalloproteinases) peptide motifs, then printed into ∼1 cm3 droplets using a BioX6 extrusion bioprinter (Figure 1A). 1 mL of media was placed on each and they were cultured in 34°C at 5% CO2 and ambient oxygen (21%) for 12 days, with 50% media changes every other day. The media consisted of Human Plasma-Like Medium (HPLM) with 15% CTS™ KnockOut™ SR XenoFree Medium, vitamins, metabolites, antioxidants, growth factors and hormones known to be present in the testis niche. Tissues were analyzed after 12 days by quantitative real-time polymerase chain reaction (qPCR) for gene expression associated with apoptosis, somatic cell function, spermatogonial stem cell maintenance, and spermatogenesis. RESULTS: After 12 days gene expression associated with somatic cell function and spermatogenesis was higher than suspension culture controls in all hydrogel groups (Figure 1B). Compared to freshly harvested testicular tissues, COL- and YIGSR-functionalized hydrogels exhibited similar expression levels of genes associated with somatic cell function, and the highest increased levels of genes associated with early spermatogenesis (Figure 1B). CONCLUSIONS: COL- or YIGSR peptide-functionalized hydrogels sustain testicular tissue function ex vivo in short-term cultures, whereas suspension culture exhibited tissue function loss, suggesting the utility of functionalized hydrogels for human IVS. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Meghan Robinson More articles by this author Ryan Flannigan More articles by this author Expand All Advertisement PDF downloadLoading ...