Improvements in Maturity and Stability of 3D iPSC-Derived Hepatocyte-like Cell Cultures
Author:
Suominen Siiri1ORCID, Hyypijev Tinja1, Venäläinen Mari1, Yrjänäinen Alma23, Vuorenpää Hanna23, Lehti-Polojärvi Mari4ORCID, Räsänen Mikko5, Seppänen Aku5, Hyttinen Jari4, Miettinen Susanna23ORCID, Aalto-Setälä Katriina16, Viiri Leena E.1
Affiliation:
1. Heart Group, Finnish Cardiovascular Research Center and Science Mimicking Life Research Center, Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, Finland 2. Adult Stem Cell Group, Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, Finland 3. Research, Development and Innovation Centre, Tampere University Hospital, 33520 Tampere, Finland 4. Computational Biophysics and Imaging Group, Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, Finland 5. Department of Technical Physics, University of Eastern Finland, 70210 Kuopio, Finland 6. Heart Hospital, Tampere University Hospital, 33520 Tampere, Finland
Abstract
Induced pluripotent stem cell (iPSC) technology enables differentiation of human hepatocytes or hepatocyte-like cells (iPSC-HLCs). Advances in 3D culturing platforms enable the development of more in vivo-like liver models that recapitulate the complex liver architecture and functionality better than traditional 2D monocultures. Moreover, within the liver, non-parenchymal cells (NPCs) are critically involved in the regulation and maintenance of hepatocyte metabolic function. Thus, models combining 3D culture and co-culturing of various cell types potentially create more functional in vitro liver models than 2D monocultures. Here, we report the establishment of 3D cultures of iPSC-HLCs alone and in co-culture with human umbilical vein endothelial cells (HUVECs) and adipose tissue-derived mesenchymal stem/stromal cells (hASCs). The 3D cultures were performed as spheroids or on microfluidic chips utilizing various biomaterials. Our results show that both 3D spheroid and on-chip culture enhance the expression of mature liver marker genes and proteins compared to 2D. Among the spheroid models, we saw the best functionality in iPSC-HLC monoculture spheroids. On the contrary, in the chip system, the multilineage model outperformed the monoculture chip model. Additionally, the optical projection tomography (OPT) and electrical impedance tomography (EIT) system revealed changes in spheroid size and electrical conductivity during spheroid culture, suggesting changes in cell–cell connections. Altogether, the present study demonstrates that iPSC-HLCs can successfully be cultured in 3D as spheroids and on microfluidic chips, and co-culturing iPSC-HLCs with NPCs enhances their functionality. These 3D in vitro liver systems are promising human-derived platforms usable in various liver-related studies, specifically when using patient-specific iPSCs.
Funder
Finnish Foundation for Cardiovascular Research Centre of Excellence in Body-on-Chip Research Competitive State Research Financing of the Expert Responsibility area of Tampere University Hospital Tampere University Doctoral School Emil Aaltonen Foundation
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