)［Abstract］ObjectiveTo study the effect of cryoprotectant Ⅰon the cryopreservation of mice hepatocytes，so as to explore a cryoprotectant with better effect for improving the cryopreservation quality of hepatocytes．MethodsKunming mice weighting 20～30g were as donors, an improved collagenase perfusion technique was established to isolate the mice hepatocytes. The isolated mice hepatocyte were cryopreserved respectively with cryoprotectant  Ⅰ( trial group) and the standard cryoprotectant ( control group) in nitrogen liquid. Cryopreserved mice hepatocytes were thawed at  0.5 month,1 month, 1.5 months, 2 months respectively.  The viability,  function and morphology of hepatocytes were observed. ResultsAfter 2 months cryopreservation,  the viability of thawed hepatocytes in trial group were (80.18±2.44)% with trypanblue staining, while the control group were (49.71±3.51)%, there was significant difference between two groups (t=23.64, P＜0.05）; the leakage rate of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) of trial group were(14.03±2.21)U/L,  (15.14±3.03)U/L, and (15.11±2.10)U/L respectively, and the control group were (24.28±1.96)U/L, (25.44±2.06)U/L and  (26.22±3.23)U/L respectively, there were significant differences between two groups respectively (t=8.84, 8.58,8.32;  all  P＜0.05）; the synthesis of albumin (ALB) of the trial group and  control group were  (3.24±0.18)g/L and  (2.56±0.33)g/L respectively, there was significant differences between two groups (t=9.25, P<0.05).There were no obvious differences in the same group of its viability, the leakage rates of ALT, AST and LDH and synthesis of ALB at each stage of thawing ( all P＞0.05). ConclusionIn this research,  compared with standard cryoprotectant, the cryoprotectant Ⅰcan effectively protect the mice hepatocytes from cryopreserved injury,  mice hepatocytes can be cryopreserved in nitrogen liquid for two months with no changes in viability.