Differentiation of the human liver progenitor cell line (HepaRG) on a microfluidic‐based biochip

Abstract

HepaRG is a bipotent stem cell line that can be differentiated towards hepatocyte-like and biliary-like cells. The entire cultivation process requires 1 month and relies on the addition of 2% dimethyl sulfoxide (DMSO) to the culture. Our motivation in this research is to differentiate HepaRG cells (progenitor cells and undifferentiated cells) towards hepatocyte-like cells by minimizing the cultivation time and without using DMSO treatment by instead using a microfluidic device combined with the following strategies: (a) comparison of extracellular matrices (matrigel and collagen I), (b) types of flow (one or both sides), and (c) effects of DMSO. Our results demonstrate that matrigel promotes the differentiation of progenitor cells towards hepatocytes and biliary-like cells. Moreover, the frequent formation of HepaRG cell clusters was observed by a supply of both sides of flow, and the cell viability and liver specific functions were influenced by DMSO. Finally, differentiated HepaRG progenitor cells cultured in a microfluidic device for 14 days without DMSO treatment yielded 70%of hepatocyte-like cells with a highly polarized organization that reacted to stimulation with IL-6 to produce C-reactive protein (CRP). This culture model has high potential for investigating cell differentiation and liver pathophysiology research.

Comprehensive Evaluation of Organotypic and Microphysiological Liver Models for Prediction of Drug-Induced Liver Injury

“…Mimetas provides microfluidic plates consisting of culture chambers with three juxtaposed inlets and outlets in which perfusion is generated by gravity on a rocking device. While this platform has been extensively used for renal and neural cultures, only a single proof-of-concept study has been presented to date, which indicated that hepatic cells (HepaRG) remained largely viable and polarized for 14 days (Jang et al, 2019).…”

Hepatic Differentiation of Stem Cells in 2D and 3D Biomaterial Systems

“…To induce differentiation to hepatocytes specifically, HPC can be cultured in the presence of HGF and FGF9 [ 39 ]. Meanwhile, human HPC cell lines, such as HepaRG, are also available for studying hepatic differentiation [ 40 ], and a similar stem cell population, termed resident liver stem cells and different from hepatic progenitor/stem cells by not expressing albumin, have also been reported [ 41 ]. One of the disadvantages, however, is the shortage of donor liver for HPC or liver stem cell isolation.…”

A Versatile Microfluidic Tool for the 3D Culture of HepaRG Cells Seeded at Various Stages of Differentiation

“…Because of their small size, liver-on-a-chip devices offer a convenient tool to culture hepatic cells under a flow and produce an adequate supply of high-throughput data for drug development 7,8 . Using such systems, cells are generally cultured as either 2D monolayers, spheroids or aggregates within an oversized chamber [9][10][11][12][13][14] . Several research groups have taken advantage of microfluidic techniques to reproduce the microarchitecture of the liver.…”