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Cells. 2019 May 31;8(6). pii: E525. doi: 10.3390/cells8060525.

Improved Oxygen Supply to Multicellular Spheroids Using A Gas-permeable Plate and Embedded Hydrogel Beads.

Author information

1
Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. n175275c@yokohama-cu.ac.jp.
2
Faculty of Science, International College of Arts and Science, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. i170239g@yokohama-cu.ac.jp.
3
Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. n175352c@yokohama-cu.ac.jp.
4
Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. n165351e@yokohama-cu.ac.jp.
5
Department of Chemical Systems Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. marie-s@chemsys.t.u-tokyo.ac.jp.
6
Department of Chemical Systems Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. masaki@chemsys.t.u-tokyo.ac.jp.
7
Department of Chemical Systems Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. sakaiyasu@chemsys.t.u-tokyo.ac.jp.
8
Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. akama@yokohama-cu.ac.jp.
9
Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. nobuhiko@yokohama-cu.ac.jp.
10
Faculty of Science, International College of Arts and Science, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan. nobuhiko@yokohama-cu.ac.jp.

Abstract

Culture systems for three-dimensional tissues, such as multicellular spheroids, are indispensable for high-throughput screening of primary or patient-derived xenograft (PDX)-expanded cancer tissues. Oxygen supply to the center of such spheroids is particularly critical for maintaining cellular functions as well as avoiding the development of a necrotic core. In this study, we evaluated two methods to enhance oxygen supply: (1) using a culture plate with a gas-permeable polydimethylsiloxane (PDMS) membrane on the bottom, and; (2) embedding hydrogel beads in the spheroids. Culturing spheroids on PDMS increased cell growth and affected glucose/lactate metabolism and CYP3A4 mRNA expression and subsequent enzyme activity. The spheroids, comprised of 5000 Hep G2 cells and 5000 20 µm-diameter hydrogel beads, did not develop a necrotic core for nine days when cultured on a gas-permeable sheet. In contrast, central necrosis in spheroids lacking hydrogel beads was observed after day 3 of culture, even when using PDMS. These results indicate that the combination of gas-permeable culture equipment and embedded hydrogel beads improves culture 3D spheroids produced from primary or PDX-expanded tumor cells.

KEYWORDS:

3D culture; gas-permeable plate; hydrogel beads; methylcellulose; multicellular spheroids

PMID:
31159231
PMCID:
PMC6627619
DOI:
10.3390/cells8060525
[Indexed for MEDLINE]
Free PMC Article

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