University of Groningen Human and rat organ slices
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چکیده
A method for the long-term storage of liver slices by cryopreservation was developed. The viability of liver slices was determined by analyzing the leakage of alanine aminotransferase, urea production, and the metabolism of testosterone. Rat liver slices were found to be optimally cryopreserved by exposure to for 30 min to 12% dimethylsulphoxide (v/v) on ice before freezing. Subsequent direct immersing in liquid nitrogen was more effective than the a cooling rate of ± 1°C/min, which reduced viability. Storage temperature at a temperature of -80°C lowered viability compared with storage at -196°C. These conditions for optimal cryopreservation were used to cryopreserve rat, rhesus monkey and cynomolgus monkey liver slices. The viability of these liver slices was maintained at: 78, 88 and 86%, respectively, when alanine aminotransferase content was measured; 85, 109 and 96%, respectively, when urea production was measured; and 116, 60 and 112%, respectively, when the metabolism of testosterone was measured. Viability was maintained for at least one month. The results show that, by using the method presented here, liver slices from these species can be stored while maintaining viabilities similar to initial values. This method will facilitate the optimal use of liver slices and reduce the number of experimental animals used.
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تاریخ انتشار 2017