Abstract:

BACKGROUND: With the application of bioartificial liver in clinic, a convenient, effective, practical protection of bioreactor loaded cell viability and function method, and the design of proprietary bioartificial liver cells with hypothermic protective solution are urgently needed.
OBJECTIVE: To verify the effect of trehalose as cryoprotectant in 4 ℃ C3A hepatocytes hypothermic preservation.
METHODS: C3A hepatocytes were preserved in different hypothermic preservation solution for 24, 48 and 72 hours at 4 ℃. The following preservation solutions were tested:1) Dullbecco's modified Eagle's medium (DMEM), 2) Lactated Ringer's solution, 3) Lactated Ringer's solution with different concentration of trehalose, 4) University of Wisconsin (UW) solution. The viability of hypothermic preserved C3A hepatocytes, index of cellular damage, oxygen-derived free radicals metabolic capacity of hypothermic preserved C3A hepatocytes after warm-reperfusion, and the apoptosis was observed by dual fluorescent staining method.
RESULTS AND CONCLUSION: The viability of hypothermic preserved C3A hepatocytes, index of cellular damage, and apoptosis were different in different groups. The indexs in Lactated Ringer's solution with different concentration of trehalose were significantly better than Lactated Ringer's solution, but inferior to UW solution (P < 0.01). Such parameters in Lactated Ringer's solution with different concentration of trehalose and UW solution were no significant difference with Lactated Ringer's solution (P > 0.05). Trehalose in C3A hepatocytes hypothermic preservation can effectively lessen hepatocytes apoptosis and depressed the extent of ischemia-reperfusion injury, thus, trehalose is the effective and important component in hypothermic preservation solution.