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Kofler, K; Ainoedhofer, H; Höllwarth, ME; Saxena, AK.
Fluorescence-activated cell sorting of PCK-26 antigen-positive cells enables selection of ovine esophageal epithelial cells with improved viability on scaffolds for esophagus tissue engineering.
Pediatr Surg Int. 2010; 26(1):97-104
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Autor/innen der Med Uni Graz:
Ainödhofer Herwig
Höllwarth Michael
Kofler Kristina
Saxena Amulya Kumar
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Abstract:
For esophagus tissue engineering, isolation and proliferation of esophageal epithelial cells (EEC) is a pre-requisite for scaffold seeding to create constructs. The aim of this study was to sort EEC expressing cytokeratin markers and their proliferative subpopulations; also, to investigate the viability of differentiated EEC subpopulations on collagen scaffolds. Ovine esophageal epithelial cells (OEECs) from sheep esophagus were analyzed using flow cytometry for pan cytokeratin (PCK-26) and proliferation cell nuclear antigen (PCNA). Using fluorescent-activated cell sorting, OEEC were separated and analyzed for PCNA expression. The OEEC subpopulations were seeded on collagen scaffolds for a week in vitro culture. Proliferation cell nuclear antigen was expressed in > 45% of OEEC isolated. In flow cytometry, 30% OEEC were PCK-26 positive which exhibited a high-proliferative capacity of 80%. PCK-26-negative OECC exhibited a low-proliferative capability of 13%. Scanning electron microscopy demonstrated organized attachment and uniform scaffold coverage in PCK-26-positive cells. Ovine esophageal epithelial cells can be divided into PCK-26-positive and negative subpopulations. PCK-26-positive OEEC constitute one-third of the isolated cells with high-proliferative capability. Seeding of PCK-26-positive OEEC on collagen scaffolds leads to uniform distribution of cells in vitro. In esophagus, tissue engineering PCK-26-positive OEEC subpopulation is important for optimal construct generation.
Find related publications in this database (using NLM MeSH Indexing)
Animals -
Cell Survival -
Disease Models, Animal -
Epithelial Cells - ultrastructure
Esophageal Diseases - pathology Esophageal Diseases - therapy
Esophagus - ultrastructure
Flow Cytometry - methods
Immunohistochemistry -
Intestinal Mucosa - ultrastructure
Keratins - immunology
Microscopy, Electron, Scanning -
Sheep -
Tissue Engineering - methods
Tissue Scaffolds -

Find related publications in this database (Keywords)
Esophageal epithelial cells
Cytokeratin
Proliferation
Sorting
Tissue engineering
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