Maintenance of high quality rat precision cut liver slices during culture to study hepatotoxic responses: Acetaminophen as a model compound
Maintenance of high quality rat precision cut liver slices during culture to study hepatotoxic responses: Acetaminophen as a model compound
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DOI：
10.1016/j.tiv.2017.05.001
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发表时间：
2017-08-01
影响因子：
3.2
通讯作者：
Hansen, Tanja
中科院分区：
文献类型：
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作者：
Granitzny, Anne;Knebel, Jan;Hansen, Tanja
精切肝切片(PCLiS)是一种很有前途的反映肝毒性反应的工具。然而，PCLiS的培养在不同的实验室之间有很大的差异，这可能会影响肝脏切片的性能，从而影响实验结果。在这项研究中，我们描述了一种易于使用的培养方法，它确保了最佳的切片活性和功能，以便为可重复性和可比性的PCLiS研究奠定基础。在24小时的培养期间，使用10个读数来评估孵育的大鼠PCLiS的质量，包括存活率(乳酸脱氢酶、天冬氨酸转氨酶和谷氨酸脱氢酶渗漏、ATP含量)和功能参数(尿素、白蛋白产生)以及组织形态和其他描述性特征(蛋白质含量、湿重、切片厚度)。最后，将PCLiS暴露于不同浓度的对乙酰氨基酚中，以评估该模型在检测肝脏毒性反应中的适用性。十分之六的读数显示出毒性效应，并显示出极好的相互关联性。三磷酸腺苷、白蛋白和组织形态测量被认为是最敏感的读数。总之，我们的结果表明，大鼠PCLiS是一种有价值的肝脏模型，特别是如果它们是在最佳标准化条件下培养的，则可用于肝毒性研究。
Precision cut liver slices (PCLiS) represent a promising tool in reflecting hepatotoxic responses. However, the culture of PCLiS varies considerably between laboratories, which can affect the performance of the liver slices and thus the experimental outcome. In this study, we describe an easily accessible culture method, which ensures optimal slice viability and functionality, in order to set the basis for reproducible and comparable PCLiS studies. The quality of the incubated rat PCLiS was assessed during a 24 h culture period using ten readouts, which covered viability (lactate dehydrogenase-, aspartate transaminase- and glutamate dehydrogenase-leakage, ATP content) and functionality parameters (urea, albumin production) as well as histomorphology and other descriptive characteristics (protein content, wet weight, slice thickness). The present culture method resulted in high quality liver slices for 24 h. Finally, PCLiS were exposed to increasing concentrations of acetaminophen to assess the suitability of the model for the detection of hepatotoxic responses. Six out of ten readouts revealed a toxic effect and showed an excellent mutual correlation. ATP, albumin and histomorphology measurements were identified as the most sensitive readouts. In conclusion, our results indicate that rat PCLiS are a valuable liver model for hepatotoxicity studies, particularly if they are cultured under optimal standardized conditions.