体外常温机械灌注对动物肝脏保存的探讨

发布时间：2018-09-19 12:34

【摘要】：目的:建立猪的肝脏体外灌注系统并对其功能进行评价。有效的延长离体肝脏在体外保存的时间并起到一定的修复作用。了解没有红细胞的培养基是否能够对猪肝脏进行体外灌注培养。方法:实验动物为巴拿马小型猪,体重在4-5kg之间,雌雄不拘。猪肝脏经过30Min缺血和30分钟冷灌注,之后猪肝脏离体并接入体外常温灌注系统并维持6小时。灌注液由标准的RPMI1640和10%小牛血清经过氧合配置而成。结果:灌注过程中肝脏持续有胆汁分泌,各项生化指标均在参考范围,肝脏组织常规病理切片未见明显结构破坏。结论:没有红细胞或者血红蛋白的灌注液是可以对猪肝脏进行体外灌注培养的。目的:研究了氧气对DCD大鼠肝脏胆道上皮细胞增殖的影响。方法:十二大鼠肝脏DCD计划分为两组。对照组大鼠肝脏离体过程中没有氧气供应的灌注。实验组大鼠肝脏离体过程中则予以充分氧合的灌注液灌注。之后所有肝脏被连接到一个体外常温机械灌注系统,灌注液中加入5-溴脱氧尿嘧啶核苷(Brd U),作为细胞增殖的标志。经过6小时体外常温机械灌注,通过比较两组胆管上皮形态、病理和Brd U复制情况进行评估。结果:我们发现实验组的大约4.5%的胆道上皮细胞出现增殖,只有而对照组只有0.4%细胞出现阳性增殖细胞。且两组标本在细胞病理形态结构上无显著变化。结论:因此,我们的结果表明,氧气供应所需的维护在肝移植胆道上皮细胞的生理功能和显示氧气供应肝脏隔离和非原位肝脏器官培养可以提高在肝移植中胆道上皮细胞损伤的修复。
[Abstract]:Objective: to establish an extracorporeal perfusion system for porcine liver and evaluate its function. It can effectively prolong the preservation time of the isolated liver in vitro and play a certain role in repairing the liver. To determine whether the culture medium without red blood cells can be perfused with porcine liver in vitro. Methods: the experimental animal was a Panama miniature pig, weighing between 4-5kg and male and female. The porcine liver was subjected to 30Min ischemia and 30 minutes cold perfusion, then the porcine liver was isolated and connected to the in vitro normothermic perfusion system and maintained for 6 hours. The perfusate was prepared by oxygenation of standard RPMI1640 and 10% calf serum. Results: bile secretion was sustained in the liver during perfusion, all biochemical indexes were within the reference range, and no obvious structural damage was found in the routine pathological sections of the liver. Conclusion: perfusion solution without erythrocyte or hemoglobin can be used to culture porcine liver in vitro. Aim: to study the effect of oxygen on the proliferation of hepatic biliary epithelial cells in DCD rats. Methods: twelve rats with liver DCD program were divided into two groups. In the control group, there was no oxygen supply perfusion in the rat liver in vitro. In the experimental group, the liver was perfused with oxygenated perfusion. All livers were then connected to an in vitro mechanical perfusion system with 5-bromodeoxyuridine (5-bromodeoxyuridine) (Brd U), as a marker of cell proliferation. After 6 hours of mechanical perfusion at room temperature in vitro, the morphology, pathology and Brd U replication of bile duct epithelium were compared between the two groups. Results: about 4.5% of the bile duct epithelial cells in the experimental group were proliferated, while only 0.4% of the control group had positive proliferative cells. There was no significant change in the morphological structure of the cells in the two groups. Conclusion: therefore, our results show that The maintenance of oxygen supply is required in the physiological function of biliary epithelial cells in liver transplantation, and it is shown that liver isolation and non-in-situ liver organ culture can improve the repair of bile duct epithelial cell injury in liver transplantation.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R657.3

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