title: Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing.
abstract: Three dimensional multicellular aggregate, also referred to as cell spheroid or microtissue, is an indispensable tool for in vitro evaluating antitumor activity and drug efficacy. Compared with classical cellular monolayer, multicellular tumor spheroid (MCTS) offers a more rational platform to predict in vivo drug efficacy and toxicity. Nevertheless, traditional processing methods such as plastic dish culture with nonadhesive surfaces are regularly time-consuming, laborious and difficult to provide uniform-sized spheroids, thus causing poor reproducibility of experimental data and impeding high-throughput drug screening. In order to provide a robust and effective platform for in vitro drug evaluation, we present an agarose scaffold prepared with the template containing uniform-sized micro-wells in commercially available cell culture plates. The agarose scaffold allows for good adjustment of MCTS size and large-scale production of MCTS. Transparent agarose scaffold also allows for monitoring of spheroid formation under an optical microscopy. The formation of MCTS from MCF-7 cells was prepared using different-size-well templates and systematically investigated in terms of spheroid growth curve, circularity, and cell viability. The doxorubicin cytotoxicity against MCF-7 spheroid and MCF-7 monolayer cells was compared. The drug penetration behavior, cell cycle distribution, cell apoptosis, and gene expression were also evaluated in MCF-7 spheroid. The findings of this study indicate that, compared with cellular monolayer, MCTS provides a valuable platform for the assessment of therapeutic candidates in an in vivo-mimic microenvironment, and thus has great potential for use in drug discovery and tumor biology research.
authors.affiliation: Laboratory of Oral Biomedical Science and Translational Medicine, Department of Prosthodontics, School of Stomatology, Tongji University, Shanghai, P.R. China.
authors.forename: Xue
authors.initials: X
authors.lastname: Gong
authors.affiliation: Institute for Biomedical Engineering & Nano Science, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China.
authors.forename: Chao
authors.initials: C
authors.lastname: Lin
authors.affiliation: Institute for Biomedical Engineering & Nano Science, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Laboratory of Oral Biomedical Science and Translational Medicine, Department of Prosthodontics, School of Stomatology, Tongji University, Shanghai, P.R. China.
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Su
authors.affiliation: Laboratory of Oral Biomedical Science and Translational Medicine, Department of Prosthodontics, School of Stomatology, Tongji University, Shanghai, P.R. China.
authors.forename: Hang
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Laboratory of Oral Biomedical Science and Translational Medicine, Department of Prosthodontics, School of Stomatology, Tongji University, Shanghai, P.R. China.
authors.forename: Tianlin
authors.initials: T
authors.lastname: Liu
authors.affiliation: Institute for Biomedical Engineering & Nano Science, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China; Institute for Engineering and Medicine, Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Wen
authors.affiliation: Institute for Biomedical Engineering & Nano Science, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhao
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology; D055808:Drug Discovery / Q000379:methods; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D018874:Spheroids, Cellular / Q000187:drug effects*; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D004317:Doxorubicin