Cryopreservation of rat and human liver slices by rapid freezing

被引:31
作者
Day, SH [1 ]
Nicoll-Griffith, DA [1 ]
Silva, JM [1 ]
机构
[1] Merck Frosst Ctr Therapeut Res, Pointe Claire, PQ H9R 4P8, Canada
关键词
liver slices; cryopreservation; vitrification; 7-ethoxycoumarin;
D O I
10.1006/cryo.1999.2161
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The cryopreservation of human li,;er slices is a promising way to enhance the ability to test the metabolism of drug candidates. This study demonstrates the use of a novel technique for the cryopreservation of both rat and human liver slices. In this technique the slices are treated with Me2SO and sandwiched between aluminum plates separated by a thin gasket. The device is then submerged in liquid nitrogen to freeze the slices. which can then be stored until use, To thaw the slices, the apparatus is submerged in a water bath at 37 degrees C. Slices frozen and thawed in this manner were compared to those frozen in conventional cryovials. The viability of the slices was determined by incubating them in 12-well plates and measuring urea synthesis. ethoxycoumarin metabolism and cytosolic enzyme leakage (LDH and ALT). The viability of mt slices frozen between plates approached that of fresh slices and was consistently higher than slices frozen in cryovials. Slices from two human samples gave similar results. The technique was found to work over a wide range of Me2SO concentrations (4.5 to 22% was tested) with an optimal concentration between 10 and 15%. (C) 1999 Academic Press.
引用
收藏
页码:154 / 159
页数:6
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