生物人工肝用C3A与L-02肝细胞零下非结冰保存后的生物学特性比较  

Comparison of biological features of C3A and L-02 hepatocytes undergoing subzero nonfreezing storage for use in bioartificial liver support system

作　　者：沙杭[1] 李庆勇[1] 张清华[1] 蒋知新[1] 李安全[1] 林虎[1] 高德禄[1] 张鋆歆[1] 高毅[2] 

机构地区：[1]中国人民解放军305医院老年病中心,北京市100017 [2]南方医科大学珠江医院肝胆二科,广东省广州市510262

出　　处：《世界华人消化杂志》2010年第33期3559-3564,共6页World Chinese Journal of Digestology

基　　金：国家高技术研究发展计划(863计划)基金资助项目;No.2006AA02A141;全军医学科研"十一五"计划专项基金资助项目;No.08Z017~~

摘　　要：目的:比较UW液零下非结冰(-0.8℃)保存后C3A与L-02细胞的生物学特性,探索其对生物人工肝的应用价值.方法:贴壁培养C3A与L-02细胞,0.25%胰酶消化,制备成细胞悬液,细胞浓度调至2×109/L,分装至2mL冻存管,UW液-0.8℃保存0、24、48及72h后,分别测定细胞存活率与凋亡率、AST与LDH释放、尿素合成功能及白蛋白分泌功能.结果:随着低温保存时间的延长,细胞存活率逐渐下降,但72hC3A细胞存活率明显高于L-02细胞(86.49%±2.80%vs70.17%±2.82%,P<0.001);细胞凋亡率亦逐渐上升,但72hC3A细胞凋亡率低于L-02细胞(1.26%±0.84%vs5.73%±1.68%,P<0.001);AST及LDH的释放量逐渐增加,但72hC3A细胞低于L-02细胞(4.61U/L±0.43U/Lvs6.22U/L±0.65U/L,P<0.001;86.10U/L±11.10U/Lvs113.88U/L±5.64U/L,P<0.001).尿素合成功能呈下降的趋势,但72hL-02细胞明显优于C3A细胞(1.01mmol/L±0.14mmol/Lvs0.60mmol/L±0.06mmol/L,P<0.001).白蛋白分泌功能逐渐降低,但72hC3A细胞明显优于L-02细胞(2.060g/L±0.220g/Lvs9.04mg/L±0.53mg/L,P<0.001).结论:UW液-0.8℃保存C3A与L-02细胞72h仍然可以满足生物人工肝支持系统的需要.以L-02细胞为材料的人工肝可能更适用于肝衰竭合并肝性脑病,以C3A细胞为材料的人工肝可能更适用于肝衰竭合并低白蛋白血症.AIM： To evaluate the biological features of C3A and L-02 hepatocytes undergoing subzero nonfreezing storage in University of Wisconsin （UW） solution for use in bioartificial liver support systems （BALSS） METHODS： C3A and L-02 hepatocytes were cultured, digested with 0.25% trypsin, prepared into a cell suspension, adjusted to a density of 2 x 10^6/mL, and stored in UW solution. After 0, 24, 48 and 72 h of hypothermic storage （-0.8 °C）, cell viability and apoptosis, LDH and AST release, and urea synthesis and albumin secretion were measured. RESULTS： Hypothermic storage decreased cell viability, urea synthesis and albumin secretion, but increased apoptosis and AST and LDH release in both C3A and L-02 hepatocytes. After 72 h of hypothermic storage, C3A hepatocytes had higher viability （86.49% ± 2.80% vs 70.17% ±2.82%, P 〈 0.001） and albumin secretion （2 060 mg/L ± 220 mg/L vs 9.04 mg/L ± 0.53 rag/L, P 〈 0.001） but lower apoptosis rate （1.26% ＋ 0.84% vs 5.73% ± 1.68%, P 〈 0.001）, LDH and AST re- lease （4.61 U/L ± 0.43 U/L vs 6.22 U/L ±0.65 U/L, P 〈 0.001; 86.10 U/L ± 11.10 U/L vs 113.88 U/L ± 5.64 U/L, P 〈 0.001） and urea synthesis （1.01 mmol/L ＋ 0.14 mmol/L vs 0.60 mmol/L 〉0.06 mmol/L, P 〈 0.001） than L-02 hepatocytes. CONCLUSION： Both C3A and L-02 hepatocytes could be stored for more than 72 h in UW solu- tion. C3A hepatocytes are more probably suitable for BALSS for management of liver failure with hepatic encephalopathy, while L-02 hepatocytes are more probably suitable for BALSS for management of liver failure with low albuminemia.

关 键 词：生物人工肝 零下非结冰 细胞凋亡 

分 类 号：R318.14[医药卫生—生物医学工程]