de Kanter Ruben, Tuin Annemarie, van de Kerkhof Esther, Martignoni Marcella, Draaisma Annelies L, de Jager Marina H, de Graaf Inge A M, Meijer Dirk K F, Groothuis Geny M M
Nerviano Medical Science Srl., Viale Pasteur 10, 20014 Nerviano (MI), Italy.
J Pharmacol Toxicol Methods. 2005 Jan-Feb;51(1):65-72. doi: 10.1016/j.vascn.2004.07.007.
A new technique was developed to prepare precision-cut slices from small intestine and colon with the object of studying the biotransformation of drugs in these organs.
Rat intestinal slices were prepared in two different ways. In the first method, slices were punched out of the small intestine. In the second method, precision-cut slices were made from agarose-filled and -embedded intestines, using the Krumdieck tissue slicer. This method was also applied to colon tissue. Viability of the slices was determined by analysis of intracellular ATP and RNA levels and morphology. Drug metabolizing activity was studied using lidocaine, testosterone, and 7-ethoxycoumarin (7-EC) as phase I substrates, and 7-hydroxycoumarin (7-HC) as a phase II substrate.
Precision-cut slices made from agarose-filled and -embedded intestine better preserved ATP levels than tissue that was punched out of the intestinal wall. After 24 h of incubation, morphology in precision cut-slices showed was quite well preserved while punched out tissue was almost completely autolytic after incubation. In addition, total RNA amount and quality was much better maintained in precision-cut slices, when compared to punched out tissue. Both intestinal slices and punched-out tissue showed high, and comparable, phase I and phase II biotransformation activities.
It is concluded that preparing precision-cut 0.25 mm slices out of agarose-filled and -embedded intestine provides an improvement, compared with punched-out tissue, and that both intestinal and colon slices are useful preparations for in vitro biotransformation studies.
开发了一种新技术，用于从小肠和结肠制备精密切割切片，目的是研究这些器官中药物的生物转化。
大鼠肠切片通过两种不同方法制备。第一种方法是从小肠中冲出切片。第二种方法是使用Krumdieck组织切片机，从琼脂糖填充和包埋的肠中制备精密切割切片。该方法也应用于结肠组织。通过分析细胞内ATP和RNA水平以及形态来确定切片的活力。使用利多卡因、睾酮和7-乙氧基香豆素（7-EC）作为I相底物，以及7-羟基香豆素（7-HC）作为II相底物来研究药物代谢活性。
与从肠壁冲出的组织相比，从琼脂糖填充和包埋的肠制备的精密切割切片能更好地保存ATP水平。孵育24小时后，精密切割切片的形态保存良好，而冲出的组织在孵育后几乎完全自溶。此外，与冲出的组织相比，精密切割切片中总RNA的量和质量得到了更好的维持。肠切片和冲出的组织均显示出较高且相当的I相和II相生物转化活性。
得出的结论是，与冲出的组织相比，从琼脂糖填充和包埋的肠制备0.25毫米的精密切割切片有所改进，并且肠切片和结肠切片都是体外生物转化研究的有用制剂。
