UW液、Celsior液和HTK液低温保存生物人工肝用L-02细胞的效果比较

Comparison of L-02 hepatocyte preservation for bioartificial liver support system with UW,Celsior and HTK solution

目的比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用L-02细胞的效果。方法制备好的L-02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72 h后,分别测定细胞存活率及死亡率(流式细胞术),ALT及LDH释放,尿素合成功能及白蛋白分泌功能。结果 UW液较Celsior液和HTK液显著提高了4℃常规低温保存72 h的L-02细胞的存活率[(60.05±4.23)%vs(50.12±3.99)%、(44.20±4.67)%],均P<0.05;降低了细胞死亡率[(39.95±4.23)%vs(49.88±3.99)%、(55.80%±4.67)%],均P<0.05;抑制了ALT、LDH释放(均P<0.05);更好地维持了L-02细胞尿素合成功能[(1.03±0.23)mmol/Lvs(0.80±0.14)mmol/L、(0.48±0.05)mmol/L]和白蛋白分泌功能[(8.36±1.38)mg/Lvs(6.41±1.25)mg/L、(5.19±0.41)mg/L),均P<0.05。结论与Celsior液和HTK液相比,使用UW液4℃常规低温保存L-02肝细胞可以明显提高复温后细胞存活率,降低低温损伤引起的ALT、LDH释放,有效保护肝细胞尿素合成功能和白蛋白分泌功能,但保存时间不应超过72 h。

Objective To compare the efficacy of L -02 hepatocyte preservations with University of Wisconsin （UW） solution, Celsior （CS） solution and Histidine- tryptophan- ketoglutarate （HTK） solution at 4℃. Methods L - 02 hepatocytes were preserved in three groups for 72 h ： UW group, CS group and HTK group. Cell viability rate, cell death rate （Flow Cytometry）, alanine aminotransferase （ALT） release, lactate dehydrogenase （LDH） release, urea synthetic ability and albumin secretion were assessed for comparison after preservation. Results Significant improvements were revealed in UW group, comparing with CS or HTK groups, in cell viability （60. 05 ±4. 23）% vs （50. 12 ±3.99）% and （44. 20 ± 4. 67 ）%, respectively, P 〈 0. 05 ）, urea synthetic ability （1.03 ± 0. 23 ）mmol/L vs （0. 80 ±0. 14 ）mmol/L and （0. 48 ±0. 05 ） mmol/L, respectively, P 〈 0. 05, and albumin secretion （8.36 ± 1.38 ） mg/L vs （6.41 ± 1.25）mg/ L and （5.19 ±0. 41 ） mg/L, respectively, P 〈0. 05. Meanwhile, significant suppression cell death rate was observed in UW group comparing with those in CS and HTK groups （39. 95± 4. 23 ） % vs （49. 88 ± 3.99） % and （55.80 ± 4. 67 ） %, respectively, P 〈 0. 05, also was the release of ALT or LDH （ P 〈 0. 05 ）. Conclusion Compared with Celsior solution and Histidine - tryptophan - ketoglutarate solution, L - 02 hepatocytca stored at 4% in UW solution possesses higher cell viability rate, lower ALT and LDH release, and better urea synthetic and albumin secretion abilities.