title: A spheroid toxicity assay using magnetic 3D bioprinting and real-time mobile device-based imaging.
abstract: An ongoing challenge in biomedical research is the search for simple, yet robust assays using 3D cell cultures for toxicity screening. This study addresses that challenge with a novel spheroid assay, wherein spheroids, formed by magnetic 3D bioprinting, contract immediately as cells rearrange and compact the spheroid in relation to viability and cytoskeletal organization. Thus, spheroid size can be used as a simple metric for toxicity. The goal of this study was to validate spheroid contraction as a cytotoxic endpoint using 3T3 fibroblasts in response to 5 toxic compounds (all-trans retinoic acid, dexamethasone, doxorubicin, 5'-fluorouracil, forskolin), sodium dodecyl sulfate (+control), and penicillin-G (-control). Real-time imaging was performed with a mobile device to increase throughput and efficiency. All compounds but penicillin-G significantly slowed contraction in a dose-dependent manner (Z' = 0.88). Cells in 3D were more resistant to toxicity than cells in 2D, whose toxicity was measured by the MTT assay. Fluorescent staining and gene expression profiling of spheroids confirmed these findings. The results of this study validate spheroid contraction within this assay as an easy, biologically relevant endpoint for high-throughput compound screening in representative 3D environments.
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Hubert
authors.initials: H
authors.lastname: Tseng
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Jacob A
authors.initials: JA
authors.lastname: Gage
authors.affiliation: LC Sciences, Houston, TX 77054 USA.
authors.forename: Tsaiwei
authors.initials: T
authors.lastname: Shen
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: William L
authors.initials: WL
authors.lastname: Haisler
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Shane K
authors.initials: SK
authors.lastname: Neeley
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Sue
authors.initials: S
authors.lastname: Shiao
authors.affiliation: Department of Physics, Rice University, Houston, TX 77005 USA.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Chen
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Pujan K
authors.initials: PK
authors.lastname: Desai
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Liao
authors.affiliation: LC Sciences, Houston, TX 77054 USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Hebel
authors.affiliation: Department of Bioengineering, Rice University, Houston, TX 77005 USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Raphael
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Jeanne L
authors.initials: JL
authors.lastname: Becker
authors.affiliation: Nano3D Biosciences (n3D), Houston, TX 77030 USA.
authors.forename: Glauco R
authors.initials: GR
authors.lastname: Souza
mesh_terms: D016475:3T3 Cells; D000818:Animals; D000900:Anti-Bacterial Agents / Q000633:toxicity; D000970:Antineoplastic Agents / Q000633:toxicity*; D018929:Cell Culture Techniques; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D008280:Magnetics; D051379:Mice; D008856:Microscopy, Fluorescence; D010400:Penicillin G / Q000633:toxicity; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity; D018874:Spheroids, Cellular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D059467:Transcriptome
chemical_list: D000900:Anti-Bacterial Agents; D000970:Antineoplastic Agents; D012967:Sodium Dodecyl Sulfate; D010400:Penicillin G