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

Cells

Masaki Nishikawa¹  Marie Shinohara¹  Yasuyuki Sakai¹  Hirotaka Mihara²  Takeshi Akama²  Kanae Sayo²  Nobuhiko Kojima²  Fumiya Tao²  Mai Kugawa³ 

[1] Department of Chemical Systems Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan;Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan;Faculty of Science, International College of Arts and Science, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan;

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

DOI  :  10.3390/cells8060525

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown