摘要
目的评价一种新型去垢剂十二烷基醚硫酸钠(SLES)经主支气管途径用于制备脱细胞肺支架的效果,并自制简易生物反应器来评价种植细胞的效果,探索构建组织工程人工肺的可行性。方法取SD大鼠肺,利用Langendorff系统经主支气管灌注0.1%SLES等溶剂制备脱细胞肺支架,在支架内种植A549细胞,生物反应器内培养3 d,评价指标包括HE染色、DAPI染色、DNA含量测定。结果经主支气管途径灌注SLES可以有效的去除肺内细胞,无残留的细胞及细胞核,DNA含量去除率达到96.27%,种植A549细胞培养3 d,细胞在支架内均匀生长,DNA含量达到正常肺的64.67%。结论新型去垢剂SLES可以制备出较为理想的脱细胞肺支架,自制的生物反应器可以有效地促进种植细胞的生长。
Objective To evaluate the function of decellularized rat lung scaffolds prepared by a novel detergent sodium lauryl ether sulfate（ SLES） given through the trachea,and the effect of cell reseeding by a simple bioreactor we designed.Methods SD rat lungs were harvested and perfused with 0.1% SLES through the trachea using Langendorff. The decellularized lung scaffolds were seeded with A549 cells and cultured in a bioreactor for 3 days.The scaffolds were evaluated via HE staining,DAPI staining and DNA quantification. Results Native lung cells were effectively removed by perfusing SLES through the trachea. There were no residual cells and cell nuclei. More than 96.27% of DNA was removed. The distribution of A549 cells in the SLES scaffolds was well-proportioned after 3 days of cultivation. DNA content of cellular scaffolds increased to 64.67% of native lungs. Conclusion The ideal decellularized lung scaffolds can be prepared by using a novel detergent SLES. The bioreactor designed by us can effectively promote the growth of seed cells.
作者
马金辉
于洁
乔叶薷
侯陈玮
陈书弘
黑飞龙
Ma Jinhui;Yu Jie;Qiao Yeru;Hou Chenwei;Chen Shuhong;Hei Feilong(Department of Cardiopulmonary Bypass, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, State Key Laboratory of Cardiovascular Disease, National Center for Cardiovascular Diseases, Beijing10037 , China)
出处
《中国体外循环杂志》
2018年第2期111-114,128,共5页
Chinese Journal of Extracorporeal Circulation
基金
国家自然基金面上项目(31370993)
北京协和医学院研究生创新基金(2017-1002-02-03)
关键词
组织工程
人工肺
去垢剂
生物反应器
脱细胞支架
Tissue engineering
Artificial lung
Detergent
Bioreactor
Decellularized scaffold
