Trehalose-containing solutions enhance preservation of cultured endothelial cells

Abstract

We have developed two 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 degrees C was determined by trypan blue exclusion and MTT assay. Trypan blue exclusion showed the viability after 48 hours of cold storage to be 49.5 +/- 4.7% (mean +/- standard error) in ET-K, 59.5 +/- 0.7% in IT-K, 29.2 +/- 2.5% in EC, and 55.3 +/- 7.6% in UW (ET-K or UW versus EC, p < 0.05; IT-K versus EC, p < 0.01). MTT assay absorbance values for cells after 48 hours of cold storage were 0.366 +/- 0.0066 (mean +/- standard error) in ET-K, 0.358 +/- 0.0044 in IT-K, 0.336 +/- 0.011 in EC, and 0.362 +/- 0.0019 in UW (ET-K or UW versus EC, p < 0.05). After 120 hours, absorbance values for cells were 0.303 +/- 0.0038 in ET-K, 0.269 +/- 0.0034 in IT-K, 0.186 +/- 0.011 in EC, and 0.265 +/- 0.0066 in UW (ET-K versus UW, p < 0.05; ET-K versus IT-K, p < 0.01; ET-K, IT-K or UW versus EC, p < 0.01). As far as the ability to preserve a murine endothelial cell line at a low temperature was concerned, the ET-K solution was superior to the UW solution, the IT-K solution and UW solution were equal, and the ET-K and IT-K solutions were superior to the EC solution.