零下非结冰保存L-02细胞的效果及与细胞凋亡的关系  

The effect and the relationship with cell apoptosis of subzero nonfreezing storage of L-02 hepatocytes used by bioartificial live support system

作　　者：李庆勇[1] 张清华[1] 蒋知新[1] 李安全[1] 夏爱祥[1] 李丽[1] 陈玲[1] 高毅[2] 

机构地区：[1]解放军305医院老年病中心,北京100017 [2]南方医科大学珠江医院

出　　处：《中国老年学杂志》2010年第12期1663-1667,共5页Chinese Journal of Gerontology

基　　金：全军医学科研"十一五"计划专项课题(No.08Z017);国家高技术研究发展计划(863计划)课题(No.2006AA02A141)

摘　　要：目的采用UW液零下非结冰(-0.8℃)保存生物人工肝用L-02细胞,与常规低温(4℃及0℃)比较,探索零下非结冰保存肝细胞的效果以及同细胞凋亡的关系。方法制备好的UW液保存的L-02细胞悬液分为3组:-0.8℃组(零下非结冰组),0℃组(0℃非结冰组),4℃组(对照组)。低温保存24h、48h及72h后,分别测定细胞存活率及凋亡率(流式细胞术)、LDH释放、尿素合成功能、白蛋白分泌功能。结果零下非结冰(-0.8℃)较常规低温(0℃及4℃)明显提高了低温保存L-02细胞的存活率〔72h:-0.8℃(70.17±2.82)%vs4℃(60.05±3.17)%〕,降低了细胞凋亡率〔72h:-0.8℃(5.73±1.68)%vs4℃(9.20±2.35)%〕。零下非结冰明显抑制了LDH的释放〔72h:-0.8℃(113.88±5.64)U/Lvs4℃(170.47±11.80)U/L〕,更好地维持了L-02细胞的尿素合成功能〔72h:-0.8℃(1.01±0.14)mmol/Lvs4℃(0.66±0.09)mmol/L〕及白蛋白分泌功能〔72h:-0.8℃(9.04±0.53)μg/mlvs4℃(7.70±0.52)μg/ml〕。结论与0℃及4℃相比,零下非结冰(-0.8℃)可明显提高L-02细胞存活率,降低低温损伤引起的细胞凋亡,有效的保护肝细胞尿素合成功能和白蛋白分泌功能。Objective To investigate the effect and the relationship with cell apoptosis of subzero nonfreezing storage （ -0. 8℃ ） compared with conventional hypothermic storage （4℃ and 0℃ ）using L-02 hepatocytes which were stored in UW solution and used by bioarti- ficial live support system. Methods L-02 hepatocytes suspended in UW solution were divided into subzero nonfreezing （ - 0. 8℃ ） , zero nonfreezing （0℃） and control groups （4℃）. After 24,48 and 72 h of hypothermie storage, the cell viability rate and cell apoptosis rate （flow cytometry）, the level of lactate dehydrogenase（LDH）, the ability of hepatoeytes to synthesize urea and secrete albumin were measured. Results Significant improvement of cell viability [72 h ： - 0. 8℃ （70. 17 ± 2. 82） % vs 4℃ （60. 05±3. 17 ） % ] was observed in subzero nonfreezing group（ -0. 8℃ ） compared with that in zero nonfreezing group（0℃） and control group （4℃）. Cell apoptosis [72 h： - 0.8℃（5.73±1.68）% vs4℃（9.20±2.35）%] andLDH [72h：-0.8℃（l13.88 ±5.64）U/Lvs4℃（170.47±11.80）U/L] weresig- nificantly suppressed and the ability of hepatocytes to synthesize urea [ 72 h ： - 0. 8℃（ 1.01±0. 14 ） mmol/L vs 4℃（0. 66 ± 0. 09 ） mmol/L ] and secrete albumin [ 72 h ： - 0. 8 ℃（9.04 ±0 . 53 ） μg/ml vs 4℃ （ 7. 70±0. 52 ） μg/ml] were also better maintained in subzero nonfreezing group. Conclusions Subzero nonfreezing storage（ -0. 8℃） of L-02 hepatocytes stored in UW solution could pride better cell viability, the ability of hepatocytes to synthesize urea and secrete albumin and lower cell apoptosis. Subzero nonfreezing storage（ -0. 8℃） of hepatoeytes and constructing a ＂ready to use＂ hepatocytes bank which likes the blood bank will efficiently promote the development of the BLASS.

关 键 词：生物人工肝 零下非结冰 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]