Open AccessDifferentiation of respiratory epithelium in a 3-dimensional co-culture with fibroblasts embedded in fibrin gel
Author(s) -
Stefan Jockenhoevel,
Stefanie Albers,
Anja Lena Thiebes,
Kai L. Gessenich,
Christian Cornelissen
Publication year - 2016
Publication title -
multidisciplinary respiratory medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 28
eISSN - 2049-6958
pISSN - 1828-695X
DOI - 10.4081/mrm.2016.270
Subject(s) - cytokeratin , epithelium , fibrin , respiratory epithelium , pathology , immunohistochemistry , biology , tissue engineering , respiratory system , microbiology and biotechnology , chemistry , immunology , anatomy , medicine , genetics
Background: 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. Methods: 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. Results: 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. Conclusions: This study indicates that adding fibroblasts to fibrin gel positively influences the differentiation of respiratory epithelium.