Besiedelung und Differenzierung von respiratorischen Epithelzellen auf Fibrin Gel in statischer Kultur und verschiedenen Tiermodellen

• Cultivation and differentiation of respiratory epithelial cells onto fibrin gel in static culture conditions in different animal models

Albers, Stefanie; Jockenhövel, Stefan (Thesis advisor); Dreher, Michael (Thesis advisor)

Aachen (2018, 2019)
Dissertation / PhD Thesis

Dissertation, Rheinisch-Westfälische Technische Hochschule Aachen, 2018

Abstract

Tracheal obstruction caused by malignancies are critical situations in clinic. Untill now nobody has found any fully satisfying therapeutic options. A problem of air way stents in the clinic is the absensce of so called mucociliary clearance which is imporant to transport mucus and particles out of the lung. In tissue engineered stents this property is highly depend onto a good differentiation of the used respiratory epithelium. In this thesis the improvement of seeding a fibrin gel with respiratory epithelial cells was investigated, as well as the differentiation of primary respiratory epithelial cells with cell culture methods like co culturing with fibroblasts, culture media, air-lift interface culturen and coating of culture dishes. The time until confluence decreases with higher concentration of cells. High concentrations of cells lead to shedding of the cells and to an unequal distribution of the cells. Low concentration of cells lead to a higher contamination of respiratory epithelial cells with fibroblasts and to a longer time until confluence. Co-culturing of porcine fibroblasts in fibrin gel at a concentration of 10 *105 cells per cm³ leads to formation of a pseudostratified respiratory epithelium with the common cell types found in respiratory epithelium in vivo. Higher concentrations of fibroblasts do not have a positive effect onto the differentiation. It was not possible to differentiate respiratory epithelial cells from pig and sheep onto a collagen surface alone. The tested culture media had different effects onto the differentiation of the cells which where species dependend. In conclusion different protocols for culturing ovine and porcine respiratory epithelial cells have been established. It was possible to build a 3-dimensional co-culturing system onto a fibrin gel basis. The exact mechanisms of differentiation of respiratory epithelial cells still need further investigation.