A DMSO-free hepatocyte maturation medium accelerates hepatic differentiation of HepaRG cells in vitro.
A DMSO-free hepatocyte maturation medium accelerates hepatic differentiation of HepaRG cells in vitro.
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DOI：
10.1016/j.biopha.2019.109010
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发表时间：
2019-06
期刊：
影响因子：
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通讯作者：
Zhen-yu Wang;Wei-Jian Li;Qigen Li;Hong-Shu Jing;Tianjie Yuan;Gongbo Fu;Dan Tang;Hongdan Zhang;Hexin Yan;B. Zhai
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文献类型：
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作者：
Zhen-yu Wang;Wei-Jian Li;Qigen Li;Hong-Shu Jing;Tianjie Yuan;Gongbo Fu;Dan Tang;Hongdan Zhang;Hexin Yan;B. Zhai
研究药物肝毒性、肝病和生物人工肝的最基本工具一直是能够在体外再现肝脏生理的模型。肝祖细胞系HepaRG代表了原代肝细胞的有效替代物。然而，HepaRG的分化依赖于使用高浓度二甲基亚砜（DMSO）的长期诱导，这可能会损害药物代谢的研究并限制该肝脏模型的适用性。在这里，我们提出了一种新的肝成熟培养基（HMM）的分化HepaRG，这是基于鸡尾酒的可溶性分子，模仿体内环境。我们发现，HMM可以快速（约9天）诱导HepaRG分化为极化的肝细胞，具有良好的代谢功能。此外，在三维培养条件下，肝球状体显示出与原代人肝细胞（PHH）接近的多种肝功能和毒性特征。我们的工作证明了HMM作为HepaRG的DMSO依赖性分化方案的替代方案的实用性;此外，这些结果促进了HepaRG的应用。
The most essential tools for studying drug hepatotoxicity, liver diseases, and bioartificial livers have always been models that can recapitulate liver physiology in vitro. The liver progenitor cell line HepaRG represents an effective surrogate of the primary hepatocyte. However, the differentiation of HepaRG relies on long-term induction using a high concentration of dimethyl sulfoxide (DMSO), which may compromise the research of drug metabolism and restrict the applicability of this hepatic model. Here, we present a novel hepatic maturation medium (HMM) for the differentiation of HepaRG, which is based on a cocktail of soluble molecules that mimick the in vivo environment. We showed that HMM could rapidly (about nine days) induce HepaRG differentiation into polarized hepatocytes with maturely metabolic functions. In addition, under three-dimensional culture conditions, the hepatic spheroids showed multiple liver functions and toxicity profiles close to those of primary human hepatocytes (PHH). Our work demonstrates the utility of HMM as an alternative to the DMSO-dependent differentiation protocol for HepaRG; moreover, these results facilitate the application of HepaRG.