Optimization of Albumin Secretion and Metabolic Activity of Cytochrome P450 1A1 of Human Hepatoblastoma HepG2 Cells in Multicellular Spheroids by Controlling Spheroid Size

Tomoko Nishikawa; Yutaro Tanaka; Makiya Nishikawa; Yuka Ogino; Kosuke Kusamori; Narumi Mizuno; Yuya Mizukami; Kazunori Shimizu; Satoshi Konishi; Yuki Takahashi; Yoshinobu Takakura

doi:10.1248/bpb.b16-00833

Regular Articles

Optimization of Albumin Secretion and Metabolic Activity of Cytochrome P450 1A1 of Human Hepatoblastoma HepG2 Cells in Multicellular Spheroids by Controlling Spheroid Size

Tomoko Nishikawa, Yutaro Tanaka, Makiya Nishikawa , Yuka Ogino, Kosuke Kusamori, Narumi Mizuno, Yuya Mizukami, Kazunori Shimizu, Satoshi Konishi, Yuki Takahashi, Yoshinobu Takakura

Author information

Tomoko Nishikawa
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Yutaro Tanaka
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Makiya Nishikawa Corresponding author
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Institute for Innovative NanoBio Drug Discovery and Development Graduate School of Pharmaceutical Sciences, Kyoto University
Yuka Ogino
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Kosuke Kusamori
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Narumi Mizuno
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Yuya Mizukami
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Kazunori Shimizu
Institute for Innovative NanoBio Drug Discovery and Development Graduate School of Pharmaceutical Sciences, Kyoto University
Department of Biotechnology, Graduate School of Engineering, Nagoya University
Department of Mechanical Engineering, Ritsumeikan University
Satoshi Konishi
Institute for Innovative NanoBio Drug Discovery and Development Graduate School of Pharmaceutical Sciences, Kyoto University
Department of Mechanical Engineering, Ritsumeikan University
Ritsumeikan-Global Innovation Research Organization, Ritsumeikan University
Yuki Takahashi
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Yoshinobu Takakura
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University
Institute for Innovative NanoBio Drug Discovery and Development Graduate School of Pharmaceutical Sciences, Kyoto University

Keywords: albumin, CYP1A1, human hepatoblastoma HepG2 cell, size control, spheroid

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2017 Volume 40 Issue 3 Pages 334-338

DOI https://doi.org/10.1248/bpb.b16-00833

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Abstract

Multicellular spheroids are useful as three-dimensional cell culture systems and for cell-based therapies. Their successful application requires an understanding of the consequences of spheroid size for cellular functions. In the present study, we prepared multicellular spheroids of different sizes using the human hepatoblastoma HepG2 cells, as hepatocytes are frequently used for in vitro drug screening and cell-based therapy. Precise polydimethylsiloxane-based microwells with widths of 360, 450, 560, and 770 µm were fabricated using a micromolding technique. Incubation of HepG2 cells in cell culture plates containing the microwells resulted in the formation of HepG2 spheroids with average diameters of 195, 320, 493, and 548 µm. The cell number per spheroid positively correlated with its diameter, and the viability of HepG2 cells was 94% or above for all samples. The smallest HepG2 spheroids showed the highest albumin secretion. On the other hand, the metabolic activity of 7-ethoxyresorufin, a fluorometric substrate for CYP1A1, increased with increasing spheroid size. These results indicate that controlling spheroid size is important when preparing HepG2 spheroids and that the size of HepG2 spheroids greatly influences the cellular function of HepG2 cells in the spheroids.

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