An evaluation of the utility of the hepatic differentiation method using hollow fiber/organoid culture for the development of a hybrid artificial liver device

To put the hybrid artificial liver (HAL) using cultured hepatocytes into practical use, it is necessary to develop a high-performance artificial liver device. We developed a novel hollow fiber (HF)/organoid culture method to induce the differentiation of pluripotent stem cells into hepatocytes. In this study, we compared the results of the hepatic differentiation using the HF/organoid culture with those using monolayer culture to evaluate its utility as a hepatic differentiation method. In both cell cultures, ES cells showed high proliferative activity immediately after cell seeding. The up-regulation of hepatocyte-specific markers such as albumin (ALB), carbamoyl phosphate synthetase 1 (CPS-1) and tryptophan 2,3-dioxygenase (TDO) were observed as the culture progressed, and the expression of liver-specific functions such as the removal of ammonia and albumin secretion were detected after about 2 weeks of the hepatic differentiation induction in the HF/organoid culture. However, the results were not observed in the monolayer culture. In conclusion, the HF/organoid culture method has promise as an effective tool for the differentiation of ES cells into hepatocytes.

► We developed a HF/organoid culture method to induce the differentiation of ES cells into hepatocytes. ► We compared the results of the hepatic differentiation using the HF/organoid culture with those using monolayer culture. ► The up-regulation of hepatocyte markers and the liver-specific functions were observed in the HF/organoid culture. ► However, they were not observed in the monolayer cultures. ► The HF/organoid culture method is an effective tool for inducing the differentiation of ES cells into hepatocytes.