2D/3D buccal epithelial cell self-assembling as a tool for cell phenotype maintenance and fabrication of multilayered epithelial linings in vitro. (18th July 2018)

• Record Type: Journal Article
• 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
• Is Part Of: Biomedical materials. Volume 13:Number 5(2018:Oct.)
• Publication Date: 2018-07-18
• Subjects: 2D/3D self-assembling -- 3D culture -- multilayered epithelial lining -- cell spheroids -- buccal epithelial cells -- buccal mucosa -- tissue engineered urethra; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1088/1748-605X/aace1c
• Languages: English
• ISSNs: 1748-6041

Abstract

Abstract: 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.