Korean J Thorac Cardiovasc Surg.
2008 Oct;41(5):550-562.
Study on an Effective Decellularization Technique for Cardiac Valve, Arterial Wall and Pericardium Xenographs: Optimization of Decellularization
- Affiliations
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- 1Department of Thoracic and Cardiovascular Surgery, Seoul National University Hospital, Seoul National University College of Medicine, Seoul National University Hospital Clinical Research Institute, Xenotransplantation Research Center, Korea. kyj@plaza.snu
- 2Department of Thoracic and Cardiovascular Surgery, Pusan National University Hospital, Pusan National University College of Medicine, Korea.
Abstract
- BACKGROUND: We attempted to reproduce a previously reported method that is known to be effective for decellularization, and we sought to find the optimal condition for decellularization by introducing some modificationsto this method. MATERIAL AND METHOD: Porcine semilunar valves, arterial walls and pericardium were processed for decellularization with using a variety of combinations and concentrations of decellularizing agents under different conditions of temperature, osmolarity and incubation time. The degree of decellularization and the preservation of the extracellular matrix wereevaluated by staining with hematoxylin and eosin and with alpha-Gal and DAPI in some of the decellularized tissues. RESULT: Decellularization was achieved in the specimens that were treated with sodium deoxycholate, sodium dodesyl sulfate, Triton X-100 and sodium dodesyl sulfate with Triton X-100 as single-step methods, and this was also achieved in the specimens that were treated with hypotonic solution --> Triton X-100 --> sodium dodesyl sulfate, sodium deoxycholate --> hypotonic solution --> sodium dodesyl sulfate, and hypotonic solution sodium dodesyl sulfate as multi-step methods. CONCLUSION: Considering the number and the amount of the chemicals that were used, the incubation time and the degree of damage to the extracellular matrix, a single-step method with sodium dodesyl sulfate and Triton X-100 and a multi-step method with hypotonic solution followed by sodium dodesyl sulfate were both relatively optimal methods for decellularization in this study.
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