Highly efficient SiRNA and gene transfer into hepatocyte-like HepaRG cells and primary human hepatocytes: new means for drug metabolism and toxicity studies.
Highly efficient SiRNA and gene transfer into hepatocyte-like HepaRG cells and primary human hepatocytes: new means for drug metabolism and toxicity studies.
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DOI：
10.1007/978-1-62703-321-3_25
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发表时间：
2013-01-01
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影响因子：
--
通讯作者：
Loyer, Pascal
中科院分区：
文献类型：
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作者：
Laurent, Veronique;Glaise, Denise;Loyer, Pascal
代谢能力强的肝细胞样人HepaRG细胞是替代人原代肝细胞的一种合适的体外细胞模型。在这里，我们描述了扩增祖HepaRG细胞并将其分化为肝细胞样细胞所需的培养过程。使用非病毒策略将基因和siRNA瞬时转染到培养细胞中，是破译基因功能的宝贵技术。在本章中，我们详细介绍了质粒DNA或sirna有效转移到增殖祖细胞或静止分化HepaRG细胞以及原代肝细胞的转染方案。
The metabolically competent hepatocyte-like human HepaRG cells represent a suitable alternative in vitro cell model to human primary hepatocytes. Here, we describe the culture procedure required to expand progenitor HepaRG cells and to differentiate them into hepatocyte-like cells. Transient transfection of gene and siRNA into cultured cells, using nonviral strategies, is an invaluable technique to decipher gene functions. In this chapter, we detail transfection protocols for efficient transfer of plasmid DNA or siRNAs into proliferating progenitor or quiescent differentiated HepaRG cells as well as into primary hepatocytes.