Boyden chamber-based compartmentalized tumor spheroid culture system to implement localized anticancer drug treatment

Shohei Kaneda; Jiro Kawada; Marie Shinohara; Momoko Kumemura; Ryohei Ueno; Tomoaki Kawamoto; Kenji Suzuki; Beomjoon Kim; Yoshiho Ikeuchi; Yasuyuki Sakai; Dominique Collard; Hiroyuki Fujita; Teruo Fujii

doi:10.1063/1.5125650

Boyden chamber-based compartmentalized tumor spheroid culture system to implement localized anticancer drug treatment

Biomicrofluidics. 2019 Oct 24;13(5):054111. doi: 10.1063/1.5125650. eCollection 2019 Sep.

Affiliations

¹ Center for International Research on Integrative Biomedical Systems, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 156-0041, Japan.
² LIMMS-CNRS/IIS, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 156-0041, Japan.
³ Department of Mechanical Systems Engineering, Faculty of Engineering, Kogakuin University, 2665-1 Nakano-machi, Hachioji-shi, Tokyo 192-0015, Japan.
⁴ Center for Interdisciplinary Research on Micro-Nano Methods, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 156-0041, Japan.

Abstract

In anticancer drug development, it is important to simultaneously evaluate both the effect of drugs on cell proliferation and their ability to penetrate tissues. To realize such an evaluation process, here, we present a compartmentalized tumor spheroid culture system utilizing a thin membrane with a through-hole to conduct localized anticancer treatment of tumor spheroids and monitor spheroid dimensions as an indicator of cell proliferation. The system is based on a commercialized Boyden chamber plate; a through-hole was bored through a porous membrane of the chamber, and the pre-existing 0.4 μm membrane pores were filled with parylene C. A HepG2 spheroid was immobilized onto the through-hole, separating the upper and lower compartments. Fluorescein (to verify the isolation between the compartments) and tirapazamine (TPZ; to treat only the lower part of the spheroid) were added to the upper and lower compartments, respectively. Since the transportation of fluorescein was blocked during treatment, i.e., the upper and lower compartments were isolated, it was confirmed that localized TPZ treatment was successfully conducted using the developed system. The effect of localized TPZ treatment on cell proliferation was estimated by measuring the maximum horizontal cross-sectional areas in the upper and lower parts of the spheroid by microscopic observations. This system can, thus, be used to perform localized anticancer drug treatment of tumor spheroids and evaluate the effect of drugs on cell proliferation.