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用于药物代谢和毒性研究的精密切割肝和肠切片的制备和孵育。

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

机构信息

Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands.

出版信息

Nat Protoc. 2010 Sep;5(9):1540-51. doi: 10.1038/nprot.2010.111. Epub 2010 Aug 19.

Abstract

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.

摘要

精密切割组织切片 (PCTS) 是具有可重复、定义明确厚度的组织的可行的离体外植体。它们代表了所研究器官的迷你模型,其中包含组织的所有细胞,保留了细胞间和细胞基质的相互作用的完整性,因此非常适合研究多细胞过程。PCTS 主要用于研究外源性物质的代谢和毒性,但它们也适用于许多其他用途。在这里,我们描述了制备和孵育大鼠和人肝和肠切片的方案。通过使用带有空心钻头的钻头从新鲜肝脏中制备圆柱形核心,然后使用专门设计的组织切片机从核心中切割切片。在切片之前,将肠组织嵌入琼脂糖圆柱中。当在 95% O(2)/5% CO(2) 气氛下在 6 孔或 12 孔板中孵育时,切片在 24 小时(肠)内保持活力,在 96 小时(肝)内保持活力。

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