In Vitro Culture Expansion and Characterization of Buccal Mucosal Epithelial Cells for Tissue Engineering Applications in Urethral Stricture After Transportation Using a Thermoreversible Gelation Polymer.

Introduction: The transportation of tissues from hospitals to clinical laboratories for cell therapy is an essential component of regenerative medicine. Previously, we used laboratory-cultured mucosal cells from buccal epithelium expanded and encapsulated using a scaffold-hybrid approach to the urethral stricture (BEES-HAUS) procedure. In this study, to improve the outcomes, we compared the thermoreversible gelation polymer (TGP) transportation procedure with conventional culture methods, and reported its advantages. Methods: Human buccal mucosal tissues in Phase I of the study were transported in Euro-Collins solution (ECS) and the cells obtained were cultured in two-dimensional (2D) Dulbecco's modified Eagle's medium (DMEM), CnT-Prime epithelial 2D differentiation medium (CnT-PR), and a three-dimensional (3D)-TGP scaffold. In Phase II, tissues were transported in a TGP cocktail and the ECS. The cells were cultured in 2D-DMEM and 3D-TGP, quantified, and characterized by immunohistochemistry. Results: The cells in 3D-TGP culture maintained epithelial morphology in a better manner compared with 2D-DMEM, in which they developed fibroblast-like morphology. The TGP-transported cells grew rapidly. Immunohistochemical analysis results for AE1/AE3, EGFR, integrin-β1, p63, and p75 were intensely positive in 3D-TGP. Conclusion: The TGP-based cocktail used in human buccal tissue transportation yielded cells with better morphology maintenance. The TGP scaffold provides an optimal in vitro environment wherein epithelial cells better maintain their native phenotype compared to those cultured through conventional methods. These results suggest using TGP for the transportation and culture of human buccal tissues for clinical applications. In addition, the use of a TGP-based cocktail for the transport of other tissues for regenerative medicine applications is worth further analysis.

Biopreservation and biobanking. 2021 Dec 24 [Epub ahead of print]

Akio Horiguchi, Kenichiro Ojima, Masayuki Shinchi, Yoshine Mayumi, Toshihiro Kushibiki, Shojiro Katoh, Masayuki Takeda, Masaru Iwasaki, Hiroshi Yoshioka, Vaddi Suryaprakash, Madasamy Balamurugan, Rajappa Senthilkumar, Samuel J K Abraham

Department of Urology, National Defence Medical College, Tokorozawa, Saitama, Japan., Department of Medical Engineering, National Defence Medical College, Tokorozawa, Saitama, Japan., Edogawa Evolutionary Lab of Science (EELS), Edogawa Hospital, Tokyo, Japan., Department of Urology, Faculty of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan., Center for Advancing Clinical Research (CACR), Faculty of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan., R & D Division, Mebiol Inc., Hiratsuka, Kanagawa, Japan., Department of Urology, Yashoda Hospitals, Hyderabad, Telangana, India., Department of Pathology, Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER), Karaikal, Puducherry, India., Fujio-Eiji Academic Terrain (FEAT), Nichi-In Center for Regenerative Medicine (NCRM), Chennai, Tamil Nadu, India.