Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Baumann, P.
Esophageal epithelial cell-cultures for Tissue Engineering.
[ Diplomarbeit/Master Thesis ] Graz Medical University; 2009. pp. 59
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Saxena Amulya Kumar
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- Abstract:
- Purpose: Congenital defects of the esophagus like atresia pose challenges in surgical treatment. Surgical techniques are further limited in long-gap atresia and are associated with high postoperative morbidity. In order to engineer esophageal tissue there is a requirement for 2 major cells: smooth muscle an epithelial cells. The aim of this study was to establish a reproducible protocol for explantation of esophagus, extraction and in-vitro culture of epithelial cells and demonstrate their survival on scaffolds after seeding. Methods: After esophageal explantation from Sprague-Dawley rats, the epithelial layer of was cut into 2mm pieces and placed on tissue culture treated plates. After proliferation and forming a monolayer, the cells were seeded on collagen scaffold (OptiMaix-2DTM). Histochemical analysis was performed in regular weekly intervals (week 1-8) after seeding. Results: Establishment of reproducible protocols for explantation of the esophagus, isolation and cultivation of cells, seeding on the scaffold and formation of cell-scaffold-construct in-vitro. About 48 hours after esophagus explants were attached to the Basement Membrane Matrix (BMM) (Matrigel)-precoated tissue-culture-plates the cells dissociated from the epithelium. Cell-scaffold-constructs showed cell attachment for at least 8 weeks. Purity of CK-14 positive epithelial cells was shown in regular weekly intervals by immunohistochemical and fluorescence staining. Conclusion: The established protocols serve as a basis for future work on esophageal epithelial cells, especially for in-vivo culture on scaffolds in the omentum. The results of this study will be necessary to build complex hollow organs using the hybrid approach.