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| Preparation and identification of a whole lung decellularized scaffold |
| CHEN Chan1, WANG Zhibin2, WENG Jie3, YU Qing4, XU Enpan4, CHEN Na4, MENG Zhuo4, DU Xiaohong1, CHEN Shixin2. |
1.Department of Geriatric Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 2.Department of Anatomy, Wenzhou Medical University, Wenzhou, 325035; 3.Department of Emergency, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027; 4.The Second Clinical Medical College, Wenzhou Medical University, Wenzhou, 325035.
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| Cite this article: |
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CHEN Chan,WANG Zhibin,WENG Jie, et al. Preparation and identification of a whole lung decellularized scaffold[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2016, 46(2): 88-91.
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Abstract Objective: To prepare a whole lung decellularized scaffold that was perfused with TritonX-100 and SDS, and to perform preliminary identification. Methods: Twenty SD rats were divided into two groups of 10 case in each randomly (decellularization and control group). Hearts and lungs were harvested from SD rats. In decellularization group, the arterial catheters were inserted through the aortopulmonary to establish channels for whole lung perfusion successively with heparinized PBS solution, 1% TirtonX-100, 0.8% SDS and deionized water in 37 ℃. The DNA concentration was determined after decellularization, and the scaffold and native lung were observed by HE staining, immunotluorescence and vascular cast. Results: Quantitative analysis of DNA content within the decellularized scaffold was (40.37±5.01) ng/mg, which showed a significant decrease compared to the native lung [(846.64±65.70) ng/mg]. A lot of collagen fibers could be observed with HE and immunohistochemistry stain but no visible cell nuclei remained after decellularization. Cast specimen showed that pulmonary arteries were still full and clear compared with the native lung. Conclusion: The method of perfusion with TritonX-100 and SDS can effectively remove all cellular components, and retain the extracellular matrix and vascular network structure well. It’s a convenient and ideal preparation method on decellularized lung scaffold for tissue engineering.
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Received: 15 September 2015
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