Ramboer Eva, De Craene Bram, De Kock Joey, Berx Geert, Rogiers Vera, Vanhaecke Tamara, Vinken Mathieu
In Vitro Toxicology and Dermato-Cosmetology research group, Center for Pharmaceutical Research, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussel, Belgium.
Unit of Molecular and Cellular Oncology, Inflammation Research Center, VIB, Technologiepark 927, 9052 Zwijnaarde, Belgium; Department of Biomedical Molecular Biology, Ghent University, 9052 Ghent, Belgium.
EXCLI J. 2015 Jul 28;14:875-89. doi: 10.17179/excli2015-424. eCollection 2015.
The increasing demand and hampered use of primary human hepatocytes for research purposes have urged scientists to search for alternative cell sources, such as immortalized hepatic cell lines. The aim of this study was to develop a human hepatic cell line using the combined overexpression of TERT and the cell cycle regulators cyclin D1 and mutant isoform CDK4R24C. Following transduction of adult human primary hepatocytes with the selected immortalization genes, cell growth was triggered and a cell line was established. When cultured under appropriate conditions, the cell line expressed several hepatocytic markers and liver-enriched transcription factors at the transcriptional and/or translational level, secreted liver-specific proteins and showed glycogen deposition. These results suggest that the immortalization strategy applied to primary human hepatocytes could generate a novel hepatic cell line that seems to retain some key hepatic characteristics.
用于研究目的的原代人肝细胞需求不断增加且其使用受到限制，这促使科学家寻找替代细胞来源，如永生化肝细胞系。本研究的目的是通过联合过表达端粒酶逆转录酶（TERT）以及细胞周期调节因子细胞周期蛋白D1和突变体亚型CDK4R24C来开发一种人肝细胞系。在用选定的永生化基因转导成人原代人肝细胞后，细胞生长被触发并建立了一个细胞系。在适当条件下培养时，该细胞系在转录和/或翻译水平表达多种肝细胞标志物和肝脏富集转录因子，分泌肝脏特异性蛋白质并显示糖原沉积。这些结果表明，应用于原代人肝细胞的永生化策略可以产生一种似乎保留一些关键肝脏特征的新型肝细胞系。
