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.
出处
《世界华人消化杂志》
CAS
北大核心
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~~
关键词
生物人工肝
零下非结冰
细胞凋亡
Bioartificial liver support system
Sub-zero nonfreezing
Cell apoptosis