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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 effects of UW solution, Celsior solution and HTK solution stored at 4 ℃ on L-02 cells. Methods The prepared L-02 cell suspension was divided into the following three groups: UW fluid preservation group (UW fluid group); Celsior fluid preservation group (CS fluid group); HTK fluid preservation group (HTK fluid group). Cell survival and mortality (flow cytometry), ALT and LDH release, urea synthesis and albumin secretion were measured in each group after 72 h at 4 ℃. Results Compared with Celsior solution and HTK solution, UW solution significantly increased the survival rate of L-02 cells ([(60.05 ± 4.23)% vs (50.12 ± 3.99)%, (44.20 ± 4.67)%] (P <0.05), and decreased the cell death rate [(39.95 ± 4.23)% vs (49.88 ± 3.99)%, (55.80% ± 4.67)%, P <0.05; ); Better maintain the urea synthesis function [(1.03 ± 0.23) mmol / L (0.80 ± 0.14) mmol / L, (0.48 ± 0.05) mmol / L] and albumin secretion function [(8.36 ± 1.38) mg / L vs (6.41 ± 1.25) mg / L, (5.19 ± 0.41) mg / L respectively), all P <0.05. Conclusion Compared with Celsior solution and HTK solution, conventional low temperature storage of UW solution at 4 ℃ could significantly improve cell viability after rewarming and decrease ALT and LDH release induced by hypothermia, effectively protecting liver cells from urea synthesis And albumin secretion, but the preservation time should not exceed 72 h.