Establishment of practical recellularized liver graft for blood perfusion using primary rat hepatocytes and liver sinusoidal endothelial cells
Establishment of practical recellularized liver graft for blood perfusion using primary rat hepatocytes and liver sinusoidal endothelial cells
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DOI：
10.1111/ajt.14666
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发表时间：
2018-06-01
影响因子：
8.8
通讯作者：
Uemoto, S.
中科院分区：
文献类型：
--
作者：
Kojima, H.;Yasuchika, K.;Uemoto, S.
组织脱细胞化产生了一种三维支架，可以在再细胞化后用于构建功能性肝移植。在血液灌流过程中，不适当的细胞分布和凝结阻碍了再细胞化肝脏的实际应用。本研究旨在建立一种实质细胞和内皮细胞最佳分布的接种方法，并评价肝窦状内皮细胞在去细胞肝脏中的作用。将原代大鼠肝细胞和LSECs分别经胆管和门静脉接种于脱细胞全肝支架内。胆管种植提供了适当的肝细胞分布到实质空间，门静脉种植的LSECs同时排列在门静脉腔内，从而维持了功能和形态。与单独接种的肝细胞相比，与LSECs共接种的肝细胞保留了其功能。联合种子组在体外血液灌流后，血小板沉积明显减少，肝细胞活力保持不变。总之，我们的种植方法根据去细胞肝脏的三维结构提供了最佳的细胞分布到实质和血管系统。由于其抗血栓形成作用，LSECs在工程肝的血液灌流中维持了肝功能，并支持了肝细胞的活性。这种再细胞化过程可以帮助制造出具有血液灌注性的实用肝移植物。
Tissue decellularization produces a three-dimensional scaffold that can be used to fabricate functional liver grafts following recellularization. Inappropriate cell distribution and clotting during blood perfusion hinder the practical use of recellularized livers. Here we aimed to establish a seeding method for the optimal distribution of parenchymal and endothelial cells, and to evaluate the effect of liver sinusoidal endothelial cells (LSECs) in the decellularized liver. Primary rat hepatocytes and LSECs were seeded into decellularized whole-liver scaffolds via the biliary duct and portal vein, respectively. Biliary duct seeding provided appropriate hepatocyte distribution into the parenchymal space, and portal vein-seeded LSECs simultaneously lined the portal lumen, thereby maintaining function and morphology. Hepatocytes co-seeded with LSECs retained their function compared with those seeded alone. Platelet deposition was significantly decreased and hepatocyte viability was maintained in the co-seeded group after extracorporeal blood perfusion. In conclusion, our seeding method provided optimal cell distribution into the parenchyma and vasculature according to the three-dimensional structure of the decellularized liver. LSECs maintained hepatic function, and supported hepatocyte viability under blood perfusion in the engineered liver graft owing to their antithrombogenicity. This recellularization procedure could help produce practical liver grafts with blood perfusion.