Poyck Paul P C, van Wijk Albert C W A, van der Hoeven Tessa V, de Waart Dirk R, Chamuleau Robert A F M, van Gulik Thomas M, Oude Elferink Ronald P J, Hoekstra Ruurdtje
Department of Surgery (Surgical Laboratory; IWO-1-172), Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
J Hepatol. 2008 Feb;48(2):266-75. doi: 10.1016/j.jhep.2007.09.018. Epub 2007 Dec 17.
BACKGROUND/AIMS: Clinical use of bioartificial livers (BAL) relies heavily on the development of human liver cell lines. The aim of this study was to assess the potential of the recently developed human fetal liver cell line cBAL111 for application in the AMC-BAL.
Laboratory-scale AMC-BAL bioreactors were loaded with 20 or 200 million cBAL111 cells and were cultured for 3 days. Parameters for hepatocyte-specific function and general metabolism were determined daily using tests with culture medium or 100% human serum. The bioreactors were also analyzed for mRNA levels of liver-specific genes and histology.
cBAL111 eliminated ammonia at a rate up to 49% of that in primary porcine hepatocytes (PPH), despite a low (1.1%) urea production. Transcript levels of glutamine synthetase (GS) were 570% of that in human liver, whereas genes of the urea cycle showed low expression. GS expression was confirmed immunohistochemically, and glutamine was produced by the cells. cBAL111 eliminated galactose (90.1% of PPH) and lidocaine (0.1% of PPH) and produced albumin (6% of PPH). Human serum did not increase function of cBAL111.
cBAL111 showed liver-specific functionality when cultured inside the AMC-BAL and eliminated ammonia mainly by the activity of GS, and not through the urea cycle.
背景/目的：生物人工肝（BAL）的临床应用在很大程度上依赖于人类肝细胞系的发展。本研究的目的是评估最近开发的人胎肝细胞系cBAL111在AMC - BAL中的应用潜力。
在实验室规模的AMC - BAL生物反应器中接种2000万或2亿个cBAL111细胞，并培养3天。每天使用培养基或100%人血清进行检测，以确定肝细胞特异性功能和一般代谢的参数。还对生物反应器进行了肝脏特异性基因的mRNA水平分析和组织学分析。
尽管尿素生成量较低（1.1%），但cBAL111清除氨的速率高达原代猪肝细胞（PPH）的49%。谷氨酰胺合成酶（GS）的转录水平是人肝脏中的570%，而尿素循环相关基因的表达较低。通过免疫组织化学证实了GS的表达，并且细胞产生了谷氨酰胺。cBAL111清除了半乳糖（为PPH的90.1%）和利多卡因（为PPH的0.1%），并产生了白蛋白（为PPH的6%）。人血清并未增强cBAL111的功能。
当在AMC - BAL内培养时，cBAL111表现出肝脏特异性功能，主要通过GS的活性而非尿素循环来清除氨。
