Improvements in Maturity and Stability of 3D iPSC-Derived Hepatocyte-like Cell Cultures

Siiri Suominen; Tinja Hyypijev; Mari Venäläinen; Alma Yrjänäinen; Hanna Vuorenpää; Mari Lehti-Polojärvi; Mikko Räsänen; Aku Seppänen; Jari Hyttinen; Susanna Miettinen; Katriina Aalto-Setälä; Leena E Viiri

doi:10.3390/cells12192368

Improvements in Maturity and Stability of 3D iPSC-Derived Hepatocyte-like Cell Cultures

Cells. 2023 Sep 27;12(19):2368. doi: 10.3390/cells12192368.

Authors

Siiri Suominen¹, Tinja Hyypijev¹, Mari Venäläinen¹, Alma Yrjänäinen^{2

3}, Hanna Vuorenpää^{2

3}, Mari Lehti-Polojärvi⁴, Mikko Räsänen⁵, Aku Seppänen⁵, Jari Hyttinen⁴, Susanna Miettinen^{2

3}, Katriina Aalto-Setälä^{1

6}, Leena E Viiri¹

Affiliations

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.

Keywords: 3D liver modeling; biomaterial; electrical impedance tomography (EIT); hepatocyte-like cell (HLC); induced pluripotent stem cells (iPSCs); microfluidic chip; optical projection tomography (OPT); spheroid culturing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques / methods
Endothelial Cells
Hepatocytes / metabolism
Humans
Induced Pluripotent Stem Cells* / metabolism
Liver

Grants and funding

The research leading to these results has received funding from The Finnish foundation for Cardiovascular Research, the Sohlberg Foundation, Centre of Excellence in Body-on-Chip Research (awards 326580, 336784, 336666, 326588, 312413), Competitive State Research Financing of the Expert Responsibility area of Tampere University Hospital, Tampere University Doctoral School, and Emil Aaltonen Foundation.