Ventilation-Based Decellularization System of the Lung.
Ventilation-Based Decellularization System of the Lung.
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DOI：
10.1089/biores.2016.0012
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发表时间：
2016
影响因子：
--
通讯作者：
Niklason LE
中科院分区：
文献类型：
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作者：
Tsuchiya T;Mendez J;Calle EA;Hatachi G;Doi R;Zhao L;Suematsu T;Nagayasu T;Niklason LE
对捐赠器官的需求大大超过了可用性。因此，正在开发器官捐赠的替代方案，如实验室工程器官。一种方法是使器官去细胞化，然后用选定的细胞（最好是来自器官接受者的细胞）重新播种。器官脱细胞化通常通过将清洁剂注入肝脏门静脉等血管来尝试。然而，在肺的情况下，气道提供了另一种潜在的给药途径，因为它在气管树和肺泡中的细胞和去污剂之间具有广泛的接触面积。在本研究中，我们介绍了一种新的基于通气的肺脱细胞系统，并将其疗效与通过肺动脉给予洗涤剂的普通脱细胞系统进行比较。通过肺动脉（血管组）或气管（气道组）给予500 mL 3-[（3-胆酰胺丙基）二甲基铵基]-1-丙磺酸盐（CHAPS）脱细胞溶液，对大鼠肺进行脱细胞。血管组使用20 cmH 2 O的重力压头输注CHAPS溶液。气道组在生物反应器中使用负压和呼气末正压输注洗涤剂，每次吸气体积为10 cc。病理和免疫组化结果表明，细胞外基质（ECM）的成分，包括蛋白多糖，弹性纤维，纤维连接蛋白和层粘连蛋白，在气道组比在血管组减少。Western blot分析显示，两组细胞均未检测到MHC I类抗原和β-actin。胶原蛋白测定显示，与天然肺相比，两组中的胶原蛋白保留了70%。糖胺聚糖（GAG）和DNA测定表明，GAG和DNA含量在两个脱细胞组强烈减少，但这些内容在气道组小于血管组。因此，在电子显微镜下肺泡壁较薄，在气道组中未观察到DNA残留。输注红细胞表明两组中毛细血管壁均得到保护，无血液渗漏。总之，我们描述了一种通过气道进行脱细胞的新方法，该方法代表了一种更严格的去除DNA和ECM的方法，并保留了毛细血管壁。
The demand for donated organs greatly exceeds the availability. Alternatives to organ donation, such as laboratory-engineered organs, are therefore being developed. One approach is to decellularize the organ and reseed it with selected cells, ideally from the organ recipient. Organ decellularization has typically been attempted by the administration of detergents into vessels such as the portal vein in the liver. However, in the case of the lung, the airway provides another potential administration route, because it has a wide contact area between cells and detergents in the tracheal tree and alveoli. In the present study, we introduce a novel ventilation-based decellularization system for the lung and compare its efficacy to ordinary decellularization systems administering detergent through the pulmonary artery. Rat lungs were decellularized using 500 mL of 3-[(3-cholamidopropyl) dimethylammonio]-1-Propanesulfonate (CHAPS) decellularization solution administrated through the pulmonary artery (vessel group) or through the trachea (airway group). The vessel group was infused CHAPS solution using a gravitational pressure head of 20 cmH2O. The airway group was infused with the detergent using negative pressure and positive end-expiratory pressure, for a volume 10cc with each inspiration in a bioreactor. Pathological and immunohistochemical findings indicated that components of the extracellular matrix (ECM), including proteoglycans, elastic fibers, fibronectin, and laminin, were more decreased in the airway group than in the vessel group. Western blot analysis showed that MHC class I antigen and β-actin were not detected in both decellularized groups. A collagen assay showed that collagen was 70% preserved in both groups compared to native lung. Glycosaminoglycan (GAG) and DNA assays showed that GAG and DNA contents were strongly diminished in both decellularized groups, but those contents were smaller in the airway group than in the vessel group. Accordingly, the alveolar wall was thinner on electron microscopy, and DNA remnants were not observed in the airway group. Infusion of red blood cells indicated that capillary walls were preserved without blood leakage in both groups. In conclusion, we describe a novel approach for decellularization through the airway that represents a more stringent method for both DNA and ECM removal, with capillary wall preservation.