Preprint Article Version 1 This version is not peer-reviewed

Improvement of Oxygen Supply to the Multicellular Spheroids Using a Gas-Permeable Plate and Embedded Hydrogel Beads

Version 1 : Received: 30 April 2019 / Approved: 3 May 2019 / Online: 3 May 2019 (14:20:44 CEST)

A peer-reviewed article of this Preprint also exists.

Mihara, H.; Kugawa, M.; Sayo, K.; Tao, F.; Shinohara, M.; Nishikawa, M.; Sakai, Y.; Akama, T.; Kojima, N. Improved Oxygen Supply to Multicellular Spheroids Using A Gas-permeable Plate and Embedded Hydrogel Beads. Cells 2019, 8, 525. Mihara, H.; Kugawa, M.; Sayo, K.; Tao, F.; Shinohara, M.; Nishikawa, M.; Sakai, Y.; Akama, T.; Kojima, N. Improved Oxygen Supply to Multicellular Spheroids Using A Gas-permeable Plate and Embedded Hydrogel Beads. Cells 2019, 8, 525.

Journal reference: Cells 2019, 8, 525
DOI: 10.3390/cells8060525

Abstract

Culture systems for 3-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 2 methods to enhance oxygen supply: (1) using culture plate with gas-permeable polydimethylsiloxane (PDMS) membrane at its 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 5000 Hep G2 cells and 5000 20 µm-diameter hydrogel beads did not develop a necrotic core for 9 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.

Subject Areas

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

Comments (0)

We encourage comments and feedback from a broad range of readers. See criteria for comments and our diversity statement.

Leave a public comment
Send a private comment to the author(s)
Views 0
Downloads 0
Comments 0
Metrics 0


×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.