[In vitro culture of cells from respiratory mucosa on foils of collagen, poly-L-lactide (PLLA) and poly-3-hydroxy-butyrate (PHB)].
BACKGROUND
The replacement of respiratory mucosa after surgical resections at different locations in the ENT-field (trachea, nasal septum, sinus maxillaris) seems to be essential or at least of great benefit for patients. Presently a satisfying solution for this challenge does not exist. Therefore we have analysed the growth of cells from respiratory mucosa on different matrices.


METHODS
Cell cultures are initiated mainly by growing out cells from tissue pieces but also by seeding of cells after enzymatic dissociation (sequential trypsinization) of mucosa. Cells were cultured with serum-containing and serum-free media on 3 different foils: made of collagen, of Poly-L-lactic acid (PLLA) and Poly-hydroxybutyric acid (PHB). Culturing time was on average about 4 - 5 weeks.


RESULTS
On all tested materials fibroblasts and epithelial cells have grown in principle. PLLA was the material with best properties concerning the aim of this study whereas culturing of cells on PHB only happens after surface modification by amino-functionalisation with plasma treatment. Visualisation of cells on materials of collagen could not realised by light-microscopy due to the 3-dimensional structure and the optical properties of this material. Here the cultures were analysed by cell membrane staining and by REM. We could not find a differentiation of epithelial cells with beating cilia 6 weeks after starting the culture. However 20 days after starting the culture on PLLA (good conditions for observation with light-microscope) cells with beating cilia could be observed, in our opinion resulting from dissociated cells from tissue pieces and not from proliferated cells.


CONCLUSIONS
In principle the analysed materials are useful for the culturing of cells from respiratory mucosa. However, for obtaining differentiated epithelial cells the culture conditions have to be modified.
HNO-Klinik und Poliklinik Otto Körner der Universität Rostock. juergen.ostwald@med.uni-rostock.de
J
J
Ostwald
S
S
Dommerich
Claudia
C
Nischan
B
B
Kramp
D002448:Cell Adhesion / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen; D003470:Culture Media; D017077:Culture Media, Conditioned; D004847:Epithelial Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D006885:Hydroxybutyrates; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D011091:Polyesters; D000091402:Prohibitins; D020545:Respiratory Mucosa / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D000094122:Polyhydroxybutyrates
D003094:Collagen; D003470:Culture Media; D017077:Culture Media, Conditioned; D006885:Hydroxybutyrates; D011091:Polyesters; D000091402:Prohibitins; C000717328:PHB protein, human; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C033616:poly(lactide)