Establishment and characterization of immortalized human hepatocyte cell line for applications in bioartificial livers
Establishment and characterization of immortalized human hepatocyte cell line for applications in bioartificial livers
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DOI：
10.1007/s10529-012-1025-1
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发表时间：
2012-12-01
影响因子：
2.7
通讯作者：
Li, LanJuan
中科院分区：
文献类型：
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作者：
Pan, XiaoPing;Li, JianZhou;Li, LanJuan
通过将猴病毒40大T抗原（SV 40 LT）转染原代人肝细胞，建立了永生化人肝细胞系HepLi 5。观察HepLi 5细胞的形态学和功能特性。逆转录-PCR（RT-PCR）和Western blotting检测SV 40 LT在HepLi 5细胞中的表达。RT-PCR检测肝富集基因谷氨酰胺合成酶、白蛋白和细胞色素P450的mRNA表达。检测药物代谢P450酶之一CYP 1A 2的活性。将HepLi 5细胞皮下注射到裸鼠体内，在3个月内没有诱导肿瘤。短串联重复序列结果证实了细胞系的真实性。临床级数量的HepLi 5细胞可以使用滚瓶中的大规模培养收获，之后它们的细胞功能显著增强。因此，永生化HepLi 5细胞是生物人工肝应用的合适细胞来源。
An immortalized human hepatocyte cell line, HepLi5, was established via transfection of Simian virus 40 large T antigen (SV40 LT) into primary human hepatocytes. The morphologic and functional characteristics of HepLi5 cells were evaluated. The expression of SV40 LT in HepLi5 cells was detected by reverse transcription-PCR (RT-PCR) and western blotting. mRNA expression of liver-enriched genes, including glutamine synthetase, albumin, and cytochrome P450 was detected via RT-PCR in HepLi5 cells. Activity of CYP1A2, one of the drug-metabolizing P450 enzymes, was detected. Subcutaneous injection of HepLi5 cells into nude mice did not induce tumors within 3 months. Short Tandem Repeat results confirmed the authenticity of the cell line. Clinical-grade quantities of HepLi5 cells could be harvested using large-scale culture in roller bottles after which their cellular function was significantly enhanced. Therefore, the immortalized HepLi5 cells are a suitable cell source for applications in bioartificial livers.