HepaRG cells are liver bipotent progenitors acquiring hepatocytes features when differentiated in the presence of dimethylsulfoxide (DMSO). Differentiated HepaRG (dHepaRG) are considered the best surrogate model to primary human hepatocytes (PHH) and are susceptible to several hepatotropic viruses, including Hepatitis B Virus (HBV) and Hepatitis Delta Virus (HDV) infection. Despite these advantages, HepaRG cells are not widely used for the study of these two viruses because of their long differentiation process and their rather low and variable infection rates. Here, we tested the use of a cocktail of five chemicals (5C) combined or not with DMSO to accelerate the cells’ differentiation process. We found that NTCP-mediated HDV entry and replication are similar in HepaRG cells cultivated for only 1 week with 5C and DMSO or differentiated with the regular 4-week protocol. However, even though the NTCP-mediated HBV entry process seemed similar, cccDNA and subsequent HBV replication markers were lower in HepaRG cells cultivated for 1 week with 5C and DMSO compared to the regular differentiation protocol. In conclusion, we set up a new procedure allowing fast differentiation and efficient HDV-infection of HepaRG cells and identified differential culture conditions that may allow to decipher the mechanism behind the establishment of the HBV minichromosome.

中文翻译：

---

快速分化的HepaRG细胞，可感染乙型肝炎和三角洲病毒

HepaRG细胞是在存在二甲基亚砜（DMSO）的情况下分化时获得肝细胞特征的肝双能祖细胞。差异化的HepaRG（dHepaRG）被认为是人类原代肝细胞（PHH）的最佳替代模型，并且容易感染几种肝炎病毒，包括乙型肝炎病毒（HBV）和丙型肝炎三角洲病毒（HDV）感染。尽管具有这些优点，但是HepaRG细胞由于其长的分化过程以及相当低且可变的感染率而并未广泛用于研究这两种病毒。在这里，我们测试了将五种化学物质（5C）混合使用或不使用DMSO来加速细胞分化过程的情况。我们发现，NTCP介导的HDV进入和复制在用5C和DMSO仅培养1周或用常规4周方案分化的HepaRG细胞中是相似的。但是，即使NTCP介导的HBV进入过程看起来相似，与常规分化方案相比，在5C和DMSO培养1周的HepaRG细胞中cccDNA和随后的HBV复制标记也较低。总之，我们建立了一个新的程序，可以快速分化和有效感染HepaRG细胞，并确定了不同的培养条件，可以破译HBV微型染色体建立的机制。与常规分化方案相比，在用5C和DMSO培养1周的HepaRG细胞中cccDNA和随后的HBV复制标记较低。总之，我们建立了一个新的程序，可以快速分化和有效感染HepaRG细胞，并确定了不同的培养条件，可以破译HBV微型染色体建立的机制。与常规分化方案相比，在用5C和DMSO培养1周的HepaRG细胞中cccDNA和随后的HBV复制标记较低。总之，我们建立了一个新的程序，可以快速分化和有效感染HepaRG细胞，并确定了不同的培养条件，可以破译HBV微型染色体建立的机制。