猪胆管脱细胞基质作为组织工程胆管支架的可行性

摘要

Objective:To explore a feasible method to make tissue engineering scaffold materials by using decellularized matrix of the porcine bile duct, and study the co-culture condition and compatibility between the decellularized matrix of the porcine bile duct and normal human intrahepatic bile duct epithelial cells. Methods: Trypsin, sodium dodecyl sulfate and triton were applied to remove the biliary cells in vitro, and then the morphological observation, H-E staining, VG staining, DNA determination and scanning electron microscopy were used to evaluate the digestive extent of the treated bile ducts. The cells' growth and proliferation were observed and detected after co-culturing the cells and the scaffold material. Results:The cell components of the porcine bile duct were completely removed by the sodium dodecyl sulfate and triton, and the decellularized matrix proteins were kept intact. The compatibility between the decellularized matrix of the porcine bile duct and normal human intrahepatic bile duct epithelial cells was good. Conclusion: The decellularized matrix of the porcine bile duct which was made by the sodium dodecyl sulfate and triton has a potential to be scaffold materials for bile duct tissue engineering.%目的:探讨制备猪胆管脱细胞基质作为组织工程胆管支架材料的可行方案及其与正常人胆管上皮细胞复合培养的条件和相容性.方法:采用胰蛋白酶脱细胞方法和十二烷基磺酸钠+曲拉通脱细胞方法对猪胆管进行体外消化,通过形态学观察,H-E染色,VG染色,胆管基质DNA测定和扫描电镜对2种脱细胞方法进行评价.正常人胆管上皮细胞与猪胆管脱细胞基质支架复合培养,观察和检测细胞在支架上的生长和增殖情况.结果:十二烷基磺酸钠+曲拉通脱细胞方法能彻底脱除猪胆管的细胞成分且能保持基质蛋白结构完好,猪胆管脱细胞基质与正常人胆管上皮细胞具有良好的相容性.结论:应用十二烷基磺酸钠+曲拉通脱细胞方法制备的猪胆管脱细胞基质,在细胞相容性方面有作为组织工程胆管支架材料的潜在可能.