建立转基因猪-猴异种原位全肝移植模型的初步尝试  

Preliminary attempts of establishing a transgenic pig-to-monkey orthotopic liver xenotransplantation model

作　　者：李亭[1] 李杰群[1] 李强[1] 章忠强[1] 颉斌[1] 魏红江 司中洲[1] 齐海智[1] Li Ting;Li Jiequn;Li Qiang;Zhang Zhongqiang;Xie Bin;Wei Hongjiang;Si Zhongzhou;Qi Haizhi(Department of Liver Transplantation,Transplant Medicine Research Center,Second Xiangya Hospital,Central South University,Changsha 410011,China;College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650500,China)

机构地区：[1]中南大学湘雅二医院肝脏移植科,移植医学研究中心,长沙410011 [2]云南农业大学动物医学院,昆明650500

出　　处：《中华器官移植杂志》2023年第9期549-555,共7页Chinese Journal of Organ Transplantation

基　　金：湖南省重点研发计划(2021DK2002);湖南省自然科学基金(2020JJ4804、2022JJ30816);长沙市自然科学基金(kq2014237)。

摘　　要：目的探讨稳定的猪-猴异种原位全肝移植模型制作方法,为异种肝移植的临床前研究提供良好的实验工具。方法回顾性分析中南大学湘雅二医院异种肝脏移植研究团队施行的7例猪-猴异种原位全肝移植的围手术期情况及结局。供体选用基因编辑的版纳小型猪,受体选用解剖特征、生理生化及免疫机制同人类十分接近的恒河猴。在参照临床经典原位全肝移植术式的基础上,进行猪-猴异种原位全肝移植术,并对手术过程进行改进,以减少术中出血、缩短无肝期。在胆管插管留置外引流,以观察胆汁分泌情况。术前免疫诱导采用"抗胸腺细胞球蛋白+利妥昔单抗+眼镜蛇毒因子+他克莫司";术后免疫维持采用"他克莫司+吗替麦考酚酯+甲泼尼龙",同时给予抗菌抗病毒治疗、输注凝血酶调节受体凝血功能。结果共行猪-猴异种原位全肝移植7例,在建模成熟稳定的第7例模型制作中,供体获取手术时间42 min,无热缺血时间,供体修整时间87 min,供体冷保存时间为128 min;受体手术时间123 min,无肝期27 min,肝下下腔静脉阻断38 min,手术全程出血量约10 ml。术后存活4例,术后最长存活时间为27 h。3例未存活模型中,1例为麻醉意外,2例为早期手术练习。第7例模型受体术后分泌胆汁86 ml。结论良好的供体获取、高质量的血管吻合、术中减少出血量、缩短无肝期、严格的液体管理及细致全面的监护是提高猪-猴异种肝移植模型成功率的关键,优化供体基因组合及合理免疫抑制方案是进一步实现猪-猴异种肝移植模型长期存活的关键。Objective To explore the feasibility of a stable pig-to-monkey orthotopic liver transplantation(LT)model and provide a favorable experimental tool for preclinical studies of xenotransplantation.Methods In this retrospective analysis,the authors reviewed the perioperative conditions and outcomes of 7 pig-to-monkey orthotopic liver transplants performed by a xenotransplantation research team of Second Xiangya Hospital.Gene-edited Banna miniature pigs were selected as donors and rhesus monkeys with similar anatomical characteristics,physiology,biochemistry and immune mechanism to humans were selected as recipients for pig-monkey xenogeneic orthotopic LT.Based upon classic transplantation procedures,whole liver xenogeneic orthotopic transplantation was performed.Surgical processes were modified for minimizing intraoperative hemorrhage and shortening anhepatic period.A bile duct drainage tube was implanted for observing bile secretion.ATG+anti-CD20+snake venom factor and FK-506 were utilized for immunoinduction pre-operation while tacrolimus,mycophenolate mofetil(MMF)and methyl prednisolone for postoperative immunomaintenance therapy.Antibiotics and antiviral agents were also applied and thrombin complex for improving coagulation functions.Results All procedures were successfully completed.After the stability and maturity of our model,in case No.7,donor's acquisition operative duration was 42 min without heat ischemic time,donor's trimming time 87 min,donor cold retention time 128 min,recipient's operative duration 123 min and anhepatic phase 27 min.Subhepatic inferior vena cava was occluded for 38 min.Blood loss was around 10 ml.And 4/7 models survived post-operation and the longest survival time was 27 h.Among 3 non-survival cases,the causes were anesthesia accident(n=1)and immaturity of early operation(n=2).Model No.7 had a biliary secretion volume of 86 ml post-operation.Conclusions Qualified donor acquisition,high-quality vascular anastomosis,intraoperative reduction of blood loss,shortening of anhepatic period,s

关 键 词：异种移植 肝移植 动物模型 

分 类 号：R657.3[医药卫生—外科学]