Culture of Ovine Esophageal Epithelial Cells andIn VitroEsophagus Tissue Engineering
- 1 February 2010
- journal article
- research article
- Published by Mary Ann Liebert Inc in Tissue Engineering, Part C: Methods
- Vol. 16 (1), 109-114
- https://doi.org/10.1089/ten.tec.2009.0145
Abstract
Background: Esophagus replacement presents major surgical challenges both in the pediatric and in adult patients since the various surgical techniques presently employed are associated with complications and high morbidity. Aim: The aim of this study was to establish protocols for isolation and culture of ovine esophageal epithelial cells (OEEC) and to investigate their viability on collagen scaffolds for in vitro tissue engineering. Methods: OEEC were sourced from adult Austrian mountain sheep. Briefly, the esophagus was dissected, treated with dispase to separate the epithelial layer, and further subjected to a modified explants technique to isolate OEEC. The OEEC were cultured in vitro and seeded on to unidirectional two-dimensional and three-dimensional collagen scaffolds. Results: Successful protocol was established for OEEC isolation and culture. OEEC exhibited organization and differentiation after 7 days in culture, which was complete after 18 days with the formation of a single layer sheet of differentiated cells exhibiting morphology of mature esophageal epithelium. OEEC seeded on two-dimensional collagen scaffolds demonstrated viability up to 6 weeks of in vitro culture with single layer epithelium formation after 3 weeks confirmed using pan-Cytokeratin markers. OEEC on three-dimensional scaffolds were viable for 6 weeks but did not form an epithelium sheet. Conclusion: Protocols for OEEC isolation were developed and established from adult ovine esophageal tissue. The generation of sheets of esophageal epithelium in culture and the viability of OEEC on collagen scaffolds for 6 weeks in vitro was observed. The prerequisite for esophagus tissue engineering, which is the ability to form epithelium when seeded on collagen scaffolds, was demonstrated.Keywords
This publication has 30 references indexed in Scilit:
- A subpopulation of mouse esophageal basal cells has properties of stem cells with the capacity for self-renewal and lineage specificationJCI Insight, 2008
- Intrathoracic esophageal replacement by in situ tissue-engineered esophagusThe Journal of Thoracic and Cardiovascular Surgery, 2008
- Esophageal epithelium regeneration on fibronectin grafted poly(l-lactide-co-caprolactone) (PLLC) nanofiber scaffoldBiomaterials, 2007
- Viscoelasticity of Esophageal Tissue and Application of a QLV ModelJournal of Biomechanical Engineering, 2006
- Seeding of Human Endothelial Cells on Valve Containing Aortic Mini-Roots: Development of a Seeding Device and ProcedureThe Annals of Thoracic Surgery, 2005
- The experimental replacement of a cervical esophageal segment with an artificial prosthesis with the use of collagen matrix and a silicone stentThe Journal of Thoracic and Cardiovascular Surgery, 1998
- The history of esophageal surgery: pediatric aspectsPediatric Surgery International, 1997
- Long-term effects of gastric transposition in children: A physiological studyJournal of Pediatric Surgery, 1996
- Total gastric transposition: An alternative to esophageal replacement in childrenJournal of Pediatric Surgery, 1991
- Colon interposition for esophagus in childrenJournal of Pediatric Surgery, 1985