乳猪肝细胞的生物学特性观察  

Biological characteristics of suckling porcine hepatocytes for bioartificial liver

作　　者：张世昌[1] 王英杰[1] 陈志[1] 刘涛[1] 刘俊[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2007年第24期2307-2310,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30470458);重庆市重点攻关项目(2004AC5041)~~

摘　　要：目的从细胞数量和细胞功能上探讨乳猪肝细胞用于生物人工肝的可行性。方法采用RPMI1640培养液37℃培养分离的乳猪肝细胞,观察乳猪肝细胞生长和增殖特征,检测乳猪肝细胞白蛋白合成、尿素合成、氨清除量、细胞色素P450(cytochrome P450,CYP)和葡萄糖醛酸基转移酶(UDP-glucuronyl transferases,UGT)活性。结果在7d的培养过程中,乳猪肝细胞呈典型的多角形,至少可增殖7倍以上,细胞生长曲线无明显潜伏期;乳猪肝细胞白蛋白和尿素合成量分别在130ng.10-6cells.h-1和150μmol.10-6cells.h-1以上,氨清除量和UGT活性在第3天较第1天显著下降(P<0.05),其后基本维持稳定,CYP活性逐渐下降。结论体外分离培养的乳猪肝细胞具有较强的增殖能力、合成功能和生物转化功能,能够为生物人工肝提供大量高活性的肝细胞。Objective To evaluate the applicability of suckling porcine hepatocytes to the bioartificial liver in cellular quantity and function. Methods The isolated porcine hepatocytes were cultivated by RPMI1640 medium at 37 ℃ in vitro. The characteristics of the proliferating and growing suckling porcine hepatocytes were observed. Their synthesis of albumin and urea was detected. Their clearance of ammonia and activities of cytochrome P450 （CYP） and UDP-glucuronyl transferases （UGT） were analyzed. Results The growth curve of suckling porcine hepatocytes showed an obvious exponential phase of cell growth and stationary phase. There was no marked stage of latency in the growth curve. Seven-fold proliferation of suckling porcine hepatocytes were found in the cultivation period of 7 days. The functional capacity of hepatocytes （ 106 cells） was above 130 ng/h and 150 μmol/h for albumin and urea excretion, respectively. The clearance of ammonia and the activities of UGT significantly decreased at culture days 1 and 3 （ P 〈 0. 05 ）, and maintained stably later. The activities of CYP decreased continuously duiring 7-day culture. Conclusion The suckling porcine hepatocytes have good proliferation abilities and biotransformation function, which could provide a great quantity and high activity of hepatocytes for bioartificial liver.

关 键 词：乳猪 肝细胞 生物人工肝 

分 类 号：R332[医药卫生—人体生理学] R322.47[医药卫生—基础医学]