【摘要】：目的:建立豬的肝臟體外灌注系統(tǒng)并對其功能進行評價。有效的延長離體肝臟在體外保存的時間并起到一定的修復作用。了解沒有紅細胞的培養(yǎng)基是否能夠對豬肝臟進行體外灌注培養(yǎng)。方法:實驗動物為巴拿馬小型豬,體重在4-5kg之間,雌雄不拘。豬肝臟經(jīng)過30Min缺血和30分鐘冷灌注,之后豬肝臟離體并接入體外常溫灌注系統(tǒng)并維持6小時。灌注液由標準的RPMI1640和10%小牛血清經(jīng)過氧合配置而成。結果:灌注過程中肝臟持續(xù)有膽汁分泌,各項生化指標均在參考范圍,肝臟組織常規(guī)病理切片未見明顯結構破壞。結論:沒有紅細胞或者血紅蛋白的灌注液是可以對豬肝臟進行體外灌注培養(yǎng)的。目的:研究了氧氣對DCD大鼠肝臟膽道上皮細胞增殖的影響。方法:十二大鼠肝臟DCD計劃分為兩組。對照組大鼠肝臟離體過程中沒有氧氣供應的灌注。實驗組大鼠肝臟離體過程中則予以充分氧合的灌注液灌注。之后所有肝臟被連接到一個體外常溫機械灌注系統(tǒng),灌注液中加入5-溴脫氧尿嘧啶核苷(Brd U),作為細胞增殖的標志。經(jīng)過6小時體外常溫機械灌注,通過比較兩組膽管上皮形態(tài)、病理和Brd U復制情況進行評估。結果:我們發(fā)現(xiàn)實驗組的大約4.5%的膽道上皮細胞出現(xiàn)增殖,只有而對照組只有0.4%細胞出現(xiàn)陽性增殖細胞。且兩組標本在細胞病理形態(tài)結構上無顯著變化。結論:因此,我們的結果表明,氧氣供應所需的維護在肝移植膽道上皮細胞的生理功能和顯示氧氣供應肝臟隔離和非原位肝臟器官培養(yǎng)可以提高在肝移植中膽道上皮細胞損傷的修復。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R657.3

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本文編號：2250130