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Spheroid-based three-dimensional liver-on-a-chip to investigate hepatocyte-hepatic stellate cell interactions and flow effects

Authors
Lee, Seung-ANo, Da YoonKang, EdwardJu, JongilKim, Dong-SikLee, Sang-Hoon
Issue Date
2013
Publisher
ROYAL SOC CHEMISTRY
Keywords
RAT HEPATOCYTES; NONPARENCHYMAL CELLS; GENE-EXPRESSION; COCULTURE; MEMBRANES; CULTURE; SIZE; MICROCHANNEL; ACTIVATION; GENERATION
Citation
LAB ON A CHIP, v.13, no.18, pp.3529 - 3537
Indexed
SCIE
SCOPUS
Journal Title
LAB ON A CHIP
Volume
13
Number
18
Start Page
3529
End Page
3537
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/106458
DOI
10.1039/c3lc50197c
ISSN
1473-0197
Abstract
We have developed a three-dimensional (3D) liver-on-a-chip to investigate the interaction of hepatocytes and hepatic stellate cells (HSCs) in which primary 3D hepatocyte spheroids and HSCs are co-cultured without direct cell-cell contact. Here, we show that the 3D liver chip offers substantial advantages for the formation and harvesting of spheroids. The most important feature of this liver chip is that it enables continuous flow of medium to the cells through osmotic pumping, and thus requires only minimal handling and no external power source. We also demonstrate that flow assists the formation and long-term maintenance of spheroids. Additionally, we quantitatively and qualitatively investigated the paracrine effects of HSCs, demonstrating that HSCs assist in the maintenance of hepatocyte spheroids and play an important role in the formation of tight cell-cell contacts, thereby improving liver-specific function. Spheroids derived from co-cultures exhibited improved albumin and urea secretion rates compared to mono-cultured spheroids after 9 days. Immunostaining for cytochrome P450 revealed that the enzymatic activity of spheroids co-cultured for 8 days was greater than that of mono-cultured spheroids. These results indicate that this system has the potential for further development as a unique model for studying cellular interactions or as a tool that can be incorporated into other models aimed at creating hepatic structure and prolonging hepatocyte function in culture.
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