2D/3D buccal epithelial cell self-assembling as a tool for cell phenotype maintenance and fabrication of multilayered epithelial linings in vitro

• Published in: Biomedical materials (Bristol) 2018-07, Vol.13 (5), p.054104-054104
• Main Authors: Zurina, I M, Shpichka, A I, Saburina, I N, Kosheleva, N V, Gorkun, A A, Grebenik, E A, Kuznetsova, D S, Zhang, D, Rochev, Y A, Butnaru, D V, Zharikova, T M, Istranova, E V, Zhang, Y, Istranov, L P, Timashev, P S
• Format: Article
• Language: English
• Subjects: 2D/3D self-assembling; 3D culture; Antigens, CD - metabolism; Biopsy; buccal epithelial cells; buccal mucosa; Cadherins - metabolism; Cell Adhesion; Cell Culture Techniques; Cell Proliferation; cell spheroids; Collagen - chemistry; Epithelial Cells - cytology; Epithelium - physiology; Humans; Intercellular Junctions; Microscopy, Electron, Transmission; Mouth Mucosa - cytology; multilayered epithelial lining; Phenotype; Polyesters - chemistry; Regenerative Medicine; Spheroids, Cellular; tissue engineered urethra; Tissue Engineering - methods; Urothelium - cytology; Zonula Occludens-1 Protein - metabolism
• ISSN: 1748-605X; 1748-605X
• DOI: 10.1088/1748-605X/aace1c

Maintaining the epithelial status of cells in vitro and fabrication of a multilayered epithelial lining is one of the key problems in the therapy using cell technologies. When cultured in a monolayer, epithelial cells change their phenotype from epithelial to epithelial-mesenchymal or mesenchymal that makes it difficult to obtain a sufficient number of cells in a 2D culture and to use them in tissue engineering. Here, using buccal epithelial cells from the oral mucosa, we developed a novel approach to recover and maintain the stable cell phenotype and form a multilayered epithelial lining in vitro via the 2D/3D cell self-assembling. Transitioning the cells from the monolayer to non-adhesive 3D culture conditions led to formation of self-assembling spheroids, with restoration of their epithelial characteristics after epithelial-mesenchymal transition. In 7 days, the cells within spheroids restored the apical-basal polarity, and the formation of both tight (ZO1) and adherent (E-cadherin) intercellular junctions was shown. Thus, culturing buccal epithelial cells in a 3D system allowed us to recover and durably maintain the morphological and functional characteristics of epithelial cells. The multilayered epithelial lining formation was achieved after placing spheroids for 7 days onto a hybrid matrix, which consisted of collagen layers and reinforcing poly (lactide-co-glycolide) fibers and was proven promising for replacement of the urothelium. Thus, we offer an effective technique of forming multilayered epithelial linings on carrier-matrices using cell spheroids that was not previously described elsewhere and can find a wide range of applications in tissue engineering, replacement surgery, and regenerative medicine.