Recapitulation of Human Embryonic Heartbeat to Promote Differentiation of Hepatic Endoderm to Hepatoblasts

Koki Yoshimoto; Nicolas Minier; Jiandong Yang; Satoshi Imamura; Kaylene Stocking; Janmesh Patel; Shiho Terada; Yoshikazu Hirai; Ken-Ichiro Kamei

doi:10.3389/fbioe.2020.568092

Recapitulation of Human Embryonic Heartbeat to Promote Differentiation of Hepatic Endoderm to Hepatoblasts

Front Bioeng Biotechnol. 2020 Sep 8:8:568092. doi: 10.3389/fbioe.2020.568092. eCollection 2020.

Authors

Koki Yoshimoto^{1

2

3}, Nicolas Minier¹, Jiandong Yang⁴, Satoshi Imamura¹, Kaylene Stocking^{1

5}, Janmesh Patel^{1

6}, Shiho Terada¹, Yoshikazu Hirai⁴, Ken-Ichiro Kamei^{1

7

8}

Affiliations

¹ Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan.
² Department of Biosystems Science, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.
³ Laboratory of Cellular and Molecular Biomechanics, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
⁴ Department of Micro Engineering, Kyoto University, Kyoto, Japan.
⁵ Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, United States.
⁶ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, United States.
⁷ Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang, China.
⁸ Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.

Abstract

Hepatic development requires multiple sequential physicochemical environmental changes in an embryo, and human pluripotent stem cells (hPSCs) allow for the elucidation of this embryonic developmental process. However, the current in vitro methods for hPSC-hepatic differentiation, which employ various biochemical substances, produce hPSC-derived hepatocytes with less functionality than primary hepatocytes, due to a lack of physical stimuli, such as heart beating. Here, we developed a microfluidic platform that recapitulates the beating of a human embryonic heart to improve the functionality of hepatoblasts derived from hepatic endoderm (HE) in vitro. This microfluidic platform facilitates the application of multiple mechanical stretching forces, to mimic heart beating, to cultured hepatic endoderm cells to identify the optimal stimuli. Results show that stimulated HE-derived hepatoblasts increased cytochrome P450 3A (CYP3A) metabolic activity, as well as the expression of hepatoblast functional markers (albumin, cytokeratin 19 and CYP3A7), compared to unstimulated hepatoblasts. This approach of hepatic differentiation from hPSCs with the application of mechanical stimuli will facilitate improved methods for studying human embryonic liver development, as well as accurate pharmacological testing with functional liver cells.

Keywords: heart beating; hepatic endoderm; hepatoblast; human embryonic stem cells (hESC); mechanical stimulation; microfluidic device.