三种肝保存液低温保存生物人工肝用C3A细胞的效果比较

The effect comparison of hypothermic UW,Celsior and HTK solution for the storage of C3A hepatocytes

目的比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用C3A细胞的效果。方法制备好的C3A细胞悬液分以下3组:UW液保存组(UW液组);Celsior液保存组(CS液组);HTK液保存组(HTK液组)。各组细胞于4℃低温保存72 h后,分别测定细胞存活率及死亡率(流式细胞术),丙氨酸氨基转移酶(ALT)及乳酸释放,尿素合成功能及清蛋白分泌功能。结果 UW液及Celsior液比HTK液显著提高了4℃低温保存72 h的C3A细胞的存活率(78.56%±4.67%vs 76.03%±3.53%,60.54%±3.18%,P<0.05);抑制了C3A细胞低温保存时ALT释放及乳酸的释放(P<0.05);更好地维持C3A细胞尿素合成功能[(0.52±0.11)mmol/L vs(0.51±0.06)mmol/L,(0.32±0.05)mmol/L,P<0.05]和清蛋白分泌功能[(1.79±0.26)g/L vs(1.75±0.21)g/L,(1.20±0.17)g/L,P<0.05]。UW液同Celsior液4℃低温保存C3A细胞的效果无差异。结论同HTK液相比,使用UW液或者Celsior液4℃保存C3A细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT释放和乳酸释放,有效的保护肝细胞尿素合成功能和清蛋白分泌功能。

Objective To compare the effect of University of Wisconsin solution（UW）,Celsior solution（CS）and Histidine-tryptophan-ketoglutarate solution（HTK）for the storage of C3A hepatocytes at 4 ℃.Methods C3A hepatocytes were divided into the following three groups：UW group,CS group,and HTK group.After 72 h of hypothermic storage,the cell viability and death rate（Flow Cytometry）,the alanine aminotransferase（ALT）release and lactic acid production,and the ability of hepatocytes to synthesize urea and secrete albumin were respectively measured.Results Compared with that in HTK group,the cell viability（78.56%±4.67% vs 76.03%±3.53%,60.54%±3.18%,P0.05）of C3A hepatocytes which were stored for 72 h were significantly improved in UW and CS groups.The ALT release and the lactic acid production（P0.05）were significantly suppressed in UW and CS groups.The ability of hepatocytes to synthesize urea[（0.52±0.11）mmol/L vs（0.51±0.06）mmol/L,（0.32±0.05）mmol/L,P0.05]and secrete albumin[（1.79±0.26） g/L vs（1.75±0.21）g/L,（1.20±0.17）g/L,P0.05]were better maintained in UW and CS groups.There was no difference between UW and CS group for the effect of storage of C3A hepatocytes.Conclusion Compared with HTK solution,the hypothermic storage（4 ℃）of C3A hepatocytes in UW or CS solution could obviously improve the cell viability rate,decrease the ALT release and the lactic acid production,and to protect the ability of hepatocytes to synthesize urea and secrete albumin.