北京生物医学工程

灌注法大鼠全肺脏脱细胞基质支架的建立

Whole lung extracellular matrix scaffold in rats by perfusion

作者：黑飞龙周建业罗富良吴蓓陈蒙蒙杨胜男龙村

单位：                      中国医学科学院  北京协和医学院阜外心血管病医院体外循环科（北京100037）        

关键词：组织工程；肺脏；脱细胞基质

分类号：

出版年·卷·期（页码）：2013·32·1（17-21）

摘要：

目的应用灌注法建立大鼠全肺脏脱细胞基质支架，探讨灌注法制备的肺脱细胞基质作为细胞支架构建组织工程器官的可行性。方法 12周龄Wistar大鼠20只，麻醉后完整切取肺组织。由肺动脉插入18G灌注头连接到Langendorff灌注平台，保持灌注压40cmH2O。肺脏用肝素化PBS液将残留血液冲洗干净，然后用1%脱氧胆酸钠溶液灌注90min，再用核酸酶应用液灌注30min。检测指标包括常规苏木素伊红(HE)染色观察细胞形态；Weigert弹力纤维+Von Gieson结缔组织(ET+VG)染色观察胶原纤维和弹性纤维的分布。结果进行脱细胞灌注后，大鼠肺脏逐渐呈均一的白色半透明状，显微镜观察肺组织细胞已基本消失，而胶原纤维排列较整齐，弹性纤维保存良好。结论灌注法是一种简单有效构建大鼠全肺组织基质支架的技术，1%脱氧胆酸钠溶液能够较好地脱除大鼠肺脏的细胞。

Objectives To produce whole-lung extracellular matrix(ECM) scaffold in rats by perfusion and to assess the feasibility of ECM as the cytoskeleton and tissue-engineered organ construction. Methods Lungs were harvested from twenty 12-week-old Wistar rats.Lungs were explanted，cannulated with an 18G cannula，and perfused (constant pressure，40cmH2O) through the pulmonary artery on Langendorff system with heparinized phosphate-buffered saline (PBS)，1% sodium deoxycholate in deionized water (90min) and RNase & DNase away (30min).HE staining was performed to confirm the removal of cells and Von Gieson staining wasperformed to show the integrity of collagen and elastin.Results After the decellularization process,rat lung scaffolds showed white color and looked semitransparent.The cells were removed effectively from lung tissue,while the collagen and elastin were kept intact structure in the scaffolds.Conclusions Perfusion technic is simple and effective to produce rat whole-lung ECM, and 1% sodium deoxycholate solution can better remove rat lung cells.

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