Exosomes from human urine-derived stem cells in porcine urethral scaffold construction.

Abstract

This study aims to examine the role of exosomes derived from human urine-derived stem cells (hUSCs-Exo) in the in vitro construction of a composite structure comprising hUSCs and a porcine urethral decellularized matrix. hUSCs-derived exosomes were isolated and characterized, and hUSCs were treated with varying exosome concentrations to assess migration using scratch and Transwell assays. Dio-labeled hUSCs were seeded onto porcine urethral decellularized matrices and grouped by exosome concentration (0, 50, 100 μg/mL). Cell proliferation and distribution were examined under a fluorescence inverted microscope on days 1, 3, and 7. On day 7, samples were paraffin-embedded for histological analysis of cell integration. hUSCs with mesenchymal stem cell (MSC) properties were successfully isolated, and exosomes extracted via centrifugation. hUSCs-exosome (Exo) enhanced cell migration but did not significantly affect proliferation. Dio-labeling and H&E staining confirmed hUSC presence and attachment to the urethral matrix, while CD44 immunohistochemistry confirmed the presence and attachment of hUSCs within the scaffold. Exosomes derived from hUSCs did not significantly enhance cell proliferation in the construction of the porcine urethral decellularized matrix-hUSC complex. The specific exosomal cargo responsible for these differential effects on migration versus proliferation was not examined in this study and will be the focus of future investigations.