亚低温培养对猪肝细胞形态与功能的影响  

The effects of mild hypothermic culture on cell morphology and functions of porcine hepatocytes

作　　者：黄蕾 张世昌 顾春荣[1] 刘京萍 陈黎 HUANG Lei;ZHANG Shichang;GU Chunrong;LIU Jingping;CHEN Li(Department of Laboratory Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Department of Infectious Diseases,Southwest Hospital,Third Military Medical University;Department of Obstetrics,the First Affiliated Hospital of Nanjing Medical University,China)

机构地区：[1]南京医科大学第一附属医院检验学部,江苏南京210029 [2]第三军医大学西南医院传染科 [3]南京医科大学第一附属医院产科

出　　处：《胃肠病学和肝病学杂志》2021年第7期793-798,共6页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(81671836)。

摘　　要：目的评价亚低温培养对猪肝细胞形态与功能的影响,探讨亚低温保存和运输肝细胞的可行性。方法将分离的新生猪肝细胞分别于25℃、28℃、33℃条件下培养24 h后,恢复至37℃继续培养,观察亚低温培养肝细胞的形态及LDH和AST漏出量,检测复温前后肝细胞白蛋白合成率、氨清除率和尿素合成率。结果亚低温培养24 h和复温37℃培养能较好地维持肝细胞形态,各组肝细胞LDH和AST漏出量差异无统计学意义(P>0.05)。亚低温培养24 h,25℃组肝细胞活力显著低于对照组、28℃组和33℃组,28℃组和33℃组尿素合成率、白蛋白合成率、氨清除率均显著低于对照组(P<0.05)。复温37℃后,28℃组和33℃组肝细胞活力显著高于对照组;28℃组和33℃组肝细胞活力、尿素合成率和白蛋白合成率均显著高于复温前、25℃组和对照组,28℃组肝细胞氨清除率与对照组相比,差异无统计学意义(P>0.05),33℃组的肝细胞氨清除率显著高于对照组(P<0.05)。结论亚低温,特别是28℃和33℃培养可有效保持肝细胞形态和维持肝细胞功能,有望为临床生物人工肝治疗提供一种较好的肝细胞保存和运输方法。Objective To evaluate the effects of mild hypothermic culture on cell morphology and functions of porcine hepatocytes and the feasibility of preservation and transportation of hepatocytes. Methods The isolated neonatal porcine hepatocytes were cultured at 25 ℃, 28 ℃ and 33 ℃ for 24 hours, and then returned to 37 ℃ for further culture. The morphology and the leakage of LDH and AST of hepatocytes cultured in mild hypothermia were observed. Cell viability, albumin synthesis rate, ammonia clearance rate and urea synthesis rate of hepatocytes cultured in different conditions were measured.Results Porcine hepatocytes cultured in different conditions maintained typical morphology of hepatocytes. No significant difference in the leakage of LDH and AST was observed among all groups(P>0.05). When hepatocytes were cultured in mild hypothermic condition, cell viability in the 25 ℃ group was significantly lower than that in other groups, and the urea synthesis rate, albumin synthesis rate, ammonia clearance rate in the 28 ℃ group and 33 ℃ group were significantly lower than those in control group(P<0.05). After rewarming at 37 ℃, cell viability of hepatocytes in the 28 ℃ group and 33 ℃ group was significantly higher than that in control group;cell viability, urea synthesis rate and albumin synthesis rate of hepatocytes in the 28 ℃ group and 33 ℃ group were significantly higher than those before rewarming, the 25 ℃ group and control group. There was no significant difference between the 28 ℃ group and the control group in the ammonia clearance rate of hepatocytes(P>0.05), while the ammonia clearance rate of hepatocytes in the 33 ℃ group was significantly higher than that in the control group(P<0.05).Conclusion Mild hypothermic culture, especially under 28 ℃ and 33 ℃, could maintain cell morphology and functions of hepatocytes, which could be utilized for the preservation and transportation of hepatocytes in the bioartificial liver.

关 键 词：亚低温 猪肝细胞 功能 生物人工肝 

分 类 号：R575[医药卫生—消化系统]