Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Schuschnig, A.
Comparison of 3 techniques for isolation of ovine esophageal epithelial cells using cell-sorting to investigate maximal cell counts for tissue engineering.
[ Diplomarbeit/Master Thesis ] Graz Medical University; 2009. pp.65.
[OPEN ACCESS]
FullText
- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Saxena Amulya Kumar
- Altmetrics:
- Abstract:
- Purpose: The treatment of congenital anomalies like the long gap esophageal atresia often poses a challenge for pediatric surgeons. Autologous tissue for the correction of the anomaly does not exist in sufficient quantity, and the surgical techniques used are often associated with postoperative complications and high morbidity. For the development of a neoesophagus in the context of Tissue engineering, smooth muscle cells and also esophageal epithelial cells are required. The aim of the study was to develop a protocol for the isolation of ovine esophageal epithelial cells. For viability controls of the cell culture after the isolation photo documentation was performed. Methods: The esophageal epithelial cells were sourced from adult sheep esophagus. We compared three techniques of isolation, using the Casy System Cell Counter with regard to cell numbers and viability. After the three isolation techniques EpiLife was used as culture medium in all cell cultures. Results: The EpiLife method was effective, which 1, 05 x 105 cells per millilitre were obtained along with a survival of 68%. With the enzymatic method through Trypsin only 5, 67 x 104 cells per millilitre were isolated along with a survival of 52%. The immunohistochemistry stains with the streptavidin-biotin and the fluorescence method demonstrated the correct phenotype. Conclusion: The future intention is to perform tissue engineering for replacement of esophagus in humans. To achieve this successful transposition in a big animal model is necessary. During this study a protocol for the isolation of ovine esophageal epithelial cells was developed, which provided satisfactory results with regard to cell numbers and viability.