Differentiation of respiratory epithelium in a 3-dimensional co-culture with fibroblasts embedded in fibrin gel

Stefanie Albers, Anja Lena Thiebes, Kai L. Gessenich, Stefan Jockenhoevel*, Christian G. Cornelissen

*Corresponding author for this work

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Tracheal tissue engineering is a promising option for the treatment of tracheal defects. In a previous study we proved the suitability of fibrin gel as a scaffold for tracheal tissue engineering. This study investigates whether the differentiation of respiratory epithelium can be increased by culturing epithelial cells in a three dimensional system containing fibroblasts embedded into fibrin gel.Respiratory epithelial cells were isolated from porcine trachea, seeded onto a fibrin gel and kept in air-liquid-interface culture for 33?days. Morphology as well as pan-cytokeratin, MUC5AC and claudin-1 expression of cells cultured on pure fibrin gel were compared to culture on gels containing fibroblasts.After two weeks, cells seeded on pure fibrin gel were multilayered, showed hyperproliferation and dedifferentiation. Co-cultured cells built up a pseudostratified epithelium. The differentiation and organization of epithelial structure improved with respect to time. After four weeks, morphology of the co-cultured respiratory epithelium resembled native tracheal epithelium. Immunohistochemistry showed that respiratory epithelium co-cultured with fibroblasts had an increasing similarity of pan-cytokeratin expression compared to native trachea. Cells cultured without fibroblasts differed in pan-cytokeratin expression from native trachea and did not show any improvement of differentiation. Immunohistochemical staining of MUC5AC and claudin-1 proved seeded cells being respiratory epithelial cells.This study indicates that adding fibroblasts to fibrin gel positively influences the differentiation of respiratory epithelium.
Original languageEnglish
Article number6
JournalMultidisciplinary Respiratory Medicine
Publication statusPublished - 1 Mar 2016


  • Airways
  • Fibrin gel
  • Fibroblast
  • Respiratory epithelium
  • Tissue engineering

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