肾和肺的去细胞器官支架体外灌注制备方法的优化

发布时间：2018-04-28 07:50

本文选题：体外灌注 + 去细胞化　；参考：《中国生物医学工程学报》2017年03期

【摘要】：研究采用体外灌注法制备肾和肺去细胞器官支架的最优方法。将10只成年SD雄鼠作为器官支架供体,进行去细胞器官支架制备,不同器官采用不同血液循环方法进行体外大气压灌注(基于大气压力往循环体系中注入)0.75%SDS、1%Triton X-100、0.1%H_2O_2制备器官去细胞支架,通过HE染色、DAPI荧光染色方法检测去细胞程度,通过扫描电镜观察其结构脉络网和去细胞程度,通过阿利新蓝-过碘酸雪夫氏染法(AB-PAS)、Masson染色、免疫组化染色方法检测去细胞器官支架的成分保留。结果表明,采用体外大气压灌注法制备的去细胞器官支架能有效去除细胞,且糖原、胶原、纤连蛋白(Fibronectin)、弹性蛋白(Elastin)、层粘连蛋白(Laminin)、IV型胶原(Collagen IV)等有效保留,同时支架能保持良好的结构脉络网。通过大气压注入去细胞化试剂,能有效保留良好的结构脉络网,同时达到有效的去细胞程度(去细胞程度大于99%),且营养成分能有效保留。
[Abstract]:To study the optimal method of preparing renal and lung acellular organ scaffolds by in vitro perfusion. Ten adult Sprague-Dawley rats were used as scaffolds for the preparation of acellular organ scaffolds. Different organs used different blood circulation methods for extracorporeal atmospheric pressure perfusion (based on atmospheric pressure, 0.75 was injected into the circulation system, and one Triton X-1000.1H _ 2O _ 2 was used to prepare organ removal scaffolds, and the degree of acellular cells was detected by HE staining and DAPI fluorescence staining. The structural choroid network and acellular degree were observed by scanning electron microscope. The components of the scaffold were detected by immunohistochemical staining and AB-PASA Masson staining. The results showed that acellular organ scaffolds prepared by in vitro atmospheric pressure perfusion could effectively remove cells, and glycogen, collagen, fibronectinine, elastin, lamininine IV collagen and laminin IV collagen were effectively retained. At the same time, the scaffold can maintain a good structure of the network. Through atmospheric pressure injection of decellular reagents, good structural choroid net can be effectively retained, and the effective degree of acellular (the degree of acellular is greater than 99%), and the nutrient components can be effectively retained at the same time.
【作者单位】：温州医科大学药学院;
【基金】：国家自然科学基金(81571392) 浙江省卫生高层次创新人才温州市551高层次人才创新技术项目
【分类号】：R318.08;R617

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