|Budget Amount *help
¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1995 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1994 : ¥1,800,000 (Direct Cost : ¥1,800,000)
Background and Methods : We have developed two new types of preservation solutions containing trehalose. ET-Kyoto solution (ET-K) is an extracellular type and IT-Kyoto solution (IT-K) is an intracellular type. In the present study we examined with in vitro assays the ability of ET-K,IT-K,Euro-Collins (EC) and University of Wisconsin (UW) solutions to preserve a murine endothelial cell line. The viability of cells stored in the solutions at 4ﾟC was determined by trypan blue exclusion and MTT assay.
Results : Trypan blue exclusion : The viability after 48 hours of cold storage was 49.5<plus-minus>4.7% (mean<plus-minus>S.E.) in ET-K,59.5<plus-minus>0.7% in IT-K,29.2<plus-minus>2.5% in EC, and 55.3<plus-minus>7.6% in UW (ET-K or UW vs EC,p<0.05 ; IT-K vs EC,p<0.01). MTT assay : Absorbance values for cells after 48 hours of cold storage were 0.366<plus-minus>0.0066 (mean<plus-minus>S.E.) in ET-K,0.358<plus-minus>0.0044 in IT-K,0.336<plus-minus>0.011 in EC,and 0.362<plus-minus>0.0019 in UW (ET-K or UW vs EC,p<0.05). After 120 hours, absorbance values for cells were 0.303<plus-minus>0.0038 in ET-K,0.269<plus-minus>0.0034 in IT-K,0.186<plus-minus>0.011 in EC,and 0.265<plus-minus>0.0066 in UW (ET-K vs UW,p<0.05 ; ET-K vs IT-K, p<0.01 ; ET
Conclusions : As far as the ability to preserve a murine endothelial cell line at a low temperature was concerned, the ET-K solution was puperior to the UW solutions, the IT-K solution and UW solution were equal, and the ET-K and IT-K solutions were superior to the EC solution.