Ex Vivo Hepatic Perfusion Through the Portal Vein in Mouse.

Ex Vivo Hepatic Perfusion Through the Portal Vein in Mouse.
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在小鼠中通过门静脉的体内肝灌注。

DOI:
10.3791/63154
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发表时间:
2022-03-09
影响因子:
1.2
通讯作者:
Ragavan, Mukundan
Ragavan, Mukundan
中科院分区:
综合性期刊4区
文献类型:
--
作者:
Giacalone, Anthony G.;Merritt, Matthew E.;Ragavan, Mukundan

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代谢性疾病如糖尿病、糖尿病前期、非酒精性脂肪性肝病(NAFLD)和非酒精性脂肪性肝炎(NASH)正变得越来越常见。离体肝脏灌注允许在可以严格控制的营养条件下使用核磁共振(NMR)对肝脏代谢进行全面分析。由于计算机模拟仍然是评估激素作用和药物干预效果的主要理论手段,灌注肝脏仍然是了解肝脏代谢的最有价值的试验床之一。由于这些研究指导了对肝脏生理学的基本见解,因此结果必须准确且可重复。离体肝脏灌注可重复性的最大因素是手术质量。因此,我们引入了一种有组织的和流线型的方法来进行离体小鼠肝脏灌注的背景下,在原位NMR实验。我们还描述了一个独特的应用程序,并讨论在这些研究中遇到的共同问题。总体目的是提供一个简单的指导,我们已经完善了几年,我们认为在原位核磁共振实验的背景下,在肝切除和灌注获得可重复的结果的黄金标准的技术。到磁场中心的距离以及在NMR实验期间组织无法进行干预使得我们的方法新颖。
Metabolic diseases such as diabetes, pre-diabetes, non-alcoholic fatty liver disease (NAFLD), and nonalcoholic steatohepatitis (NASH) are becoming increasingly common. Ex vivo liver perfusions allow for a comprehensive analysis of liver metabolism using nuclear magnetic resonance (NMR), in nutritional conditions that can be rigorously controlled. As in silico simulations remain a primarily theoretical means of assessing hormone actions and the effects of pharmaceutical intervention, the perfused liver remains one of the most valuable test beds for understanding hepatic metabolism. As these studies guide basic insights into hepatic physiology, results must be accurate and reproducible. The greatest factor in the reproducibility of ex vivo hepatic perfusion is the quality of surgery. Therefore, we have introduced an organized and streamlined method to perform ex vivo mouse liver perfusions in the context of in situ NMR experiments. We also describe a unique application and discuss common issues encountered in these studies. The overall purpose is to provide an uncomplicated guide to a technique we have refined over several years that we deem the golden standard for obtaining reproducible results in hepatic resections and perfusions in the context of in situ NMR experiments. The distance to the center of the field for the magnet as well as the inaccessibility of the tissue to intervention during the NMR experiment makes our methods novel.
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