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Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies

机译:精密切割的肝肠切片的制备和孵育,用于药物代谢和毒性研究

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摘要

Precision-cut tissue slices (PCTS) are viable ex vivo explants of tissue with a reproducible, well defined thickness. They represent a mini-model of the organ under study and contain all cells of the tissue in their natural environment, leaving intercellular and cell-matrix interactions intact, and are therefore highly appropriate for studying multicellular processes. PCTS are mainly used to study the metabolism and toxicity of xenobiotics, but they are suitable for many other purposes. Here we describe the protocols to prepare and incubate rat and human liver and intestinal slices. Slices are prepared from fresh liver by making a cylindrical core using a drill with a hollow bit, from which slices are cut with a specially designed tissue slicer. Intestinal tissue is embedded in cylinders of agarose before slicing. Slices remain viable for 24 h (intestine) and up to 96 h (liver) when incubated in 6- or 12-well plates under 95% O_2/5% CO_2 atmosphere.
机译:精确切割的组织切片(PCTS)是可行的离体外植体组织,具有可复制的,确定的厚度。它们代表了正在研究的器官的微型模型,并包含自然环境中组织的所有细胞,使细胞间和细胞-基质相互作用保持完整,因此非常适合研究多细胞过程。 PCTS主要用于研究异种生物的代谢和毒性,但它们也适用于许多其他目的。在这里,我们描述了准备和孵化大鼠和人类肝脏和肠片的方案。切片是用带有空心钻头的钻头制成圆柱形核心,从新鲜的肝脏中制备的,然后使用专门设计的组织切片机将其切成薄片。切片前将肠组织嵌入琼脂糖圆柱体中。当在95%O_2 / 5%CO_2气氛下的6孔板或12孔板中孵育时,切片可保持24小时(肠道)和长达96小时(肝脏)的活力。

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