Olinga P, Groen K, Hof I H, De Kanter R, Koster H J, Leeman W R, Rutten A A, Van Twillert K, Groothuis G M
Groningen Institute for Drug Studies, Department of Pharmacokinetics and Drug Delivery, University Centre of Pharmacy, University of Groningen, The Netherlands.
J Pharmacol Toxicol Methods. 1997 Oct;38(2):59-69. doi: 10.1016/s1056-8719(97)00060-9.
Precision-cut liver slices are presently used for various research objects, e.g. to study metabolism, transport, and toxicity of xenobiotics. Various incubation systems are presently employed, but a systematic comparison between these incubation systems with respect to preservation of slice function has not been performed yet. Therefore, we started a comparative study to evaluate five of these systems: the shaken flask (an Erlenmeyer in a shaking water bath), the stirred-well (24-well culture plate equipped with grids and magnetic stirrers), rocker platform (6-well culture plate with Netwell insert rocked on a platform), the roller system (dynamic organ culture rolled on an insert in a glass vial), and the 6-well shaker (6-well culture plate in a shaking water bath). The liver slices were incubated in these incubation systems for 0.5, 1.5, and 24.5 h and subsequently subjected to viability and metabolic function tests. The viability of the incubated liver slices was evaluated by: potassium content, MTT assay, energy charge, histomorphology, and LDH leakage. Their metabolic functions were studied by determination of the metabolism of lidocaine, testosterone, and antipyrine. Up to 1.5 h of incubation all five incubation systems gave similar results with respect to viability and metabolic function of the liver slices. However, after 24 h, the shaken flask, the rocker platform, and the 6-well shaker incubation systems appeared to be superior to the stirred well and the roller incubation systems.
精密肝切片目前被用于各种研究目的，例如研究外源化合物的代谢、转运和毒性。目前采用了各种孵育系统，但尚未对这些孵育系统在切片功能保存方面进行系统比较。因此，我们开展了一项比较研究，以评估其中五种系统：摇瓶（置于振荡水浴中的锥形瓶）、搅拌孔板（配备网格和磁力搅拌器的24孔培养板）、摇床平台（带有置于平台上摇动的嵌套式细胞培养小室的6孔培养板）、滚筒系统（在玻璃小瓶中的插入物上滚动的动态器官培养）以及6孔摇床（置于振荡水浴中的6孔培养板）。将肝切片在这些孵育系统中孵育0.5、1.5和24.5小时，随后进行活力和代谢功能测试。通过钾含量、MTT法、能荷、组织形态学和乳酸脱氢酶泄漏来评估孵育后肝切片的活力。通过测定利多卡因、睾酮和安替比林的代谢来研究它们的代谢功能。在孵育长达1.5小时时，所有五种孵育系统在肝切片的活力和代谢功能方面给出了相似的结果。然而，在24小时后，摇瓶、摇床平台和6孔摇床孵育系统似乎优于搅拌孔板和滚筒孵育系统。
