零下非结冰UW液、Celsior液和HTK液保存C3A细胞的效果比较

摘要

Objective To compare the effect of C3A hepatocytes which were respectively stored in University of Wisconsin solution (UW), Celsior solution (CS) and Histidine-tryptophan-ketoglutarate solution (HTK) at a subzero nonfreezing temperature ( - 0. 8℃ ).Methods C3A hepatocytes were divided into UW, CS, HTK groups. After 72 h of hypothermic storage, the cell viability rate and cell death rate (flow cytometry), the lactate dehydrogenase (LDH) release and the content of ATP, the ability of hepatocytes to synthesize urea and secrete albumin were measured. Results Compared with that in HTK group, the significant improvement of cell viability [ ( 84. 34％ ±4.25％), (83.53％ ± 3.73％) vs (75.65％ ±3.01％),P＜0.001] and the content of ATP[(5.54± 1.44)μg/106cells, (5.51 ±1.31 ) μg/106 cells vs (2. 75 ± 1.03 ) μg/106 cells,P ＜0. 001 ] of C3A hepatocytes which were stored for 72 h were observed in UW and CS groups. The LDH release were significantly suppressed in UW and CS groups. The ability of hepatocytes to synthesize urea [ (0. 63 ±0. 10)mmol/L,(0.62 ± 0.06)mmol/L vs (0.39 ± 0.04)mmol/L,P＜0.001]and secrete albumin [(1.99 ±0.38) g/L,(1.96 ±0. 24)g/L vs ( 1.50 ±0. 18)g/L, P ＜0. 05 ] were better maintained in UW and CS groups. There were not difference between UW group and CS group. Conclusions Compared with HTK solution, subzero nonfreezing storage( -0. 8℃ ) of C3A hepatocytes stored in UW solution or CS solution could pride the higher cell viability rate and the content of ATP , the lower LDH release, and the better ability of hepatocytes to synthesize urea and secrete albumin. There is not difference between UW solution and CS solution, and cold storage is better less than 72 h.%目的 比较UW液、Celsior液和HTK液零下非结冰(-0.8℃)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-0.8℃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)、LDH释放、尿素合成功能及白蛋白分泌功能.结果 UW液及Celsior液比HTK液显著提高了零下非结冰保存72 h的C3A细胞的存活率[(84.34±4.25)%,(83.53±3.73)% vs (75.65±3.01)%,P<0.001]和细胞内ATP含量[(5.54±1.44)μg/106 个细胞,(5.51±1.31)μg/106个细胞 vs (2.75±1.03)μg/106 个细胞,P<0.001];抑制了LDH释放(P<0.001);更好地维持了细胞尿素合成功能[(0.63±0.10)mmol/L,(0.62±0.06)mmol/L vs (0.39±0.04)mmol/L,P<0.001]和白蛋白分泌功能[(1.99±0.38)g/L,(1.96±0.24)g/L vs (1.50±0.18)g/L,P<0.05].UW液与Celsior液零下非结冰保存C3A细胞的效果无差异.结论 同HTK液相比,使用UW液或者Celsior液零下非结冰(-0.8℃)保存C3A细胞可以明显地提高复温后细胞存活率和细胞内ATP含量,降低低温损伤引起的LDH释放,有效地保护肝细胞尿素合成功能和白蛋白分泌功能.UW液同Celsior液零下非结冰保存C3A细胞的效果无差异,保存时间不宜超过72 h.