ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Biofabrication
This article is part of the Research TopicInnovative Biofabrication Strategies and Cell-Derived Therapies for Advancing Urethral Tissue EngineeringView all articles
Isolation and characterization of epithelial cells and fibroblasts from the human penile urethra
Provisionally accepted
David Brownell1
Elissa Elia1Félix-Antoine Pellerin1
Stéphane CHABAUD1Sébastien Larochelle1
Veronique Moulin1Alexis Laungani2,3Stéphane Bolduc1,4*- 1Centre de Recherche en Organogénèse Expérimentale/LOEX, CHU de Québec-Université Laval Research Center (Regenerative Medicine Division), Université Laval, Québec, Canada
- 2Universite de Montreal, Montreal, Canada
- 3GrS Montreal, Montréal, Canada
- 4Department of Surgery, Université Laval, Québec, Canada
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Urethral strictures and hypospadias are common urological conditions for which autologous reconstruction remains challenging. Tissue engineering offers a promising alternative, yet current strategies often rely on heterotopic cell sources, potentially limiting functional integration. Here, we report the first isolation and characterization of epithelial and stromal cells from distinct regions of the human penile urethra: the spongy urethra and the proximal and distal fossa navicularis. Cells were successfully isolated from all three regions in 12 donors, with a 100% success rate across all tested protocols. Detailed characterization was performed for 3–4 donors, assessing yield, growth parameters, immunophenotype, and progenitor preservation. We evaluated 3-, 4-, and 6-mm biopsies to determine the minimal tissue size required for clinically relevant cell yields. Multiple enzymatic protocols were compared, using thermolysin or dispase for epithelial-stromal separation, followed by collagenase ± elastase digestion for stromal cell recovery. All biopsy sizes produced sufficient cells for tissue engineering. The combination of dispase and a 4-hour collagenase/elastase digestion yielded the highest cell numbers and clonogenic potential. These findings demonstrate the feasibility of harvesting high-quality, organ-specific autologous cells from minimal urethral biopsies. In addition to their clinical potential, these cells provide a foundation for preclinical disease modelling using patient-derived pathological cells, which are currently unavailable for in vitro studies of urethral disorders.
Keywords: primary cell isolation, Penile urethra, Dispase, Thermolysin, urethral epithelial cells, Urethral fibroblasts, urethral tissue engineering
Received: 25 Sep 2025; Accepted: 24 Nov 2025.
Copyright: © 2025 Brownell, Elia, Pellerin, CHABAUD, Larochelle, Moulin, Laungani and Bolduc. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Stéphane Bolduc
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