Survey of Apoptosis After Hypothermic Storage of a Pancreatic β-Cell Line.

Abstract

Insulin-dependent diabetes mellitus is one of the leading causes of death world wide. Donor-derived pancreas and islet of Langerhans transplantation are potential cures, however, postmortem ischemia impacts islet quality. The murine βt3 cell line was used as a model to study apoptosis after hypothermic storage by comparing Unisol™ with Belzer's machine perfusion solution (BMPS) and the University of Wisconsin (UW) solution. The objective was to determine which of these solutions provided the best support for βt3 cells and which solution demonstrated the least amount of apoptotic activity. Several apoptosis markers were measured that included the translocation of phosphatidylserine, caspase activity, and the formation of DNA laddering. In addition, metabolic activity and membrane integrity were also measured. The results demonstrated that the three solutions behaved similarly during overnight cold storage at 4°C. However, Unisol was consistently better than UW solution and BMPS, demonstrating better cell viability and recovery, and lower levels of apoptotic activity when all three parameters were measured. These results demonstrated that apoptosis plays an important role in the survival of cells and tissues during cold storage. Development of solutions to help prevent or decrease the levels of apoptosis after cold storage will likely improve overall cell and tissue recovery and survival in a clinical setting.