目的总结心脏死亡器官捐献(donation after cardiac death,DCD)供体供肝获取及应用于肝移植的临床经验和可行性。方法 2011年11月至2012年9月,西安交通大学第一附属医院采用Maastricht标准或中国标准,共获取18例DCD供肝,于该院完成经典原位肝移植14例,送往其他移植中心3例,放弃1例。对18例供体与在该院完成肝移植的14例受体的临床资料进行回顾性分析,了解供肝情况、受体围手术期及随访结果。结果 18例供体中符合Maastricht标准Ⅲ类5例、Ⅴ类2例,符合中国标准Ⅲ类(即脑-心双死亡标准器官捐献,donation after brain death plus cardiac death,DBCD)11例。按规范器官获取流程取得供肝。供肝的热缺血时间为11～18min,平均为14.5min;冷缺血时间为90～600min,平均为350min。14例受体均顺利完成移植手术。其中12例受体预后良好,肝功能逐渐恢复,未出现原发性移植肝无功能、血栓形成、排斥反应,但2例出现胆道狭窄并发症,经胆道支架置入术后引流通畅;重症监护室(ICU)治疗时间平均7d,术后住院时间平均23d,病情稳定后出院。1例受体术后2d死于肝衰竭,其供体原发病为冠状动脉粥样硬化性心脏病,需给予大量多巴胺维持其血压;另1例于术后当日死于腹腔内大出血,其供体为重症哮喘、心肺复苏后死亡。12例受体者平均随访时间为6个月,总体存活率为85%,肿瘤患者尚未发现复发转移。结论 DCD可以扩大供肝来源且近期效果良好,具有可行性。实施可控型DCD捐献,严格掌握供者适应证、加强器官评估、缩短热缺血时间和冷缺血时间,是保障供肝质量的重要因素。
BACKGROUND： The potential application of decellularized liver scaffold for liver regeneration is limited by severe shortage of donor organs. Attempt of using heterograft scaffold is accompanied with high risks of zoonosis and immunological rejection. We proposed that the spleen, which procured more extensively than the liver, could be an ideal source of decellularized scaffold for liver regeneration. METHODS： After harvested from donor rat, the spleen was processed by 12-hour freezing/thawing ×2 cycles, then circulation perfusion of 0.02% trypsin and 3% Triton X-100 sequentially through the splenic artery for 32 hours in total to prepare decellularized scaffold. The structure and component characteristics of the scaffold were determined by hematoxylin and eosin and immumohistochemical staining, scanning electron microscope, DNA detection, porosity measurement, biocompatibility and cytocompatibility test. Recellularization of scaffold by 5×106 bone marrow mesenchymal stem cells（BMSCs） was carried out to preliminarily evaluate the feasibility of liver regeneration by BMSCs reseeding and differentiation in decellularized splenic scaffold.RESULTS： After decellularization, a translucent scaffold, which retained the gross shape of the spleen, was generated. Histological evaluation and residual DNA quantitation revealed the remaining of extracellular matrix without nucleus and cytoplasm residue. Immunohistochemical study proved the existence of collagens I, IV, fibronectin, laminin and elastin in decellularized splenic scaffold, which showed a similarity with decellularized liver. A scanning electron microscope presented the remaining three-dimensional porous structure of extracellular matrix and small blood vessels. The poros-ity of scaffold, aperture of 45.36±4.87 μm and pore rate of 80.14%±2.99% was suitable for cell engraftment. Subcutaneous implantation of decellularized scaffold presented good histocompatibility, and recellularization of the splenic scaffold demonstrated that BMSCs could locate and
