A comparison of a sucrose-based solution with other preservation media forcold storage of isolated hepatocytes

A sucrose-based solution has been compared with other preservation solution s (University of Wisconsin (UW) solution and Marshall's citrate solution, w ith Dulbecco's medium as control) during hypothermic preservation of isolat ed mi hepatocytes for up to 72 h. Studies on the stability of liver cells a t low temperature by exclusion of trypan blue dye and morphological appeara nce were conducted. During storage beyond 24 h, there was a clear differenc e between cells stored in Dulbecco's medium and Marshall's citrate and thos e stored in sucrose-based solution and UW solution, with the former storage groups showing many cells developing large membrane "blebs" and the latter storage groups maintaining a more typical morphology and developing only s mall membrane protrusions. Dye exclusion was higher in sucrose-based soluti on (48 h, 75 +/- 7%; 72 h, 65 +/- 6%) and UW solution (48 h, 72 +/- 5%; 72 h, 63 +/- 4%) than in Marshall's citrate (48 h, 31 +/- 5%; 72 h, 10 +/- 1%) and Dulbecco's medium (48 h, 8 +/- 2%; 72 h, 5 +/- 1%) These data suggest that sucrose-based solution should be investigated further as a less comple x alternative solution for storage of isolated hepatocytes. (C) 2000 Academ ic Press.