Luciferase assay system to monitor fibroblast growth factor signal disruption in human iPSCs

Seiya Kanno; Kashu Mizota; Yusuke Okubo; Tatsuto Kageyama; Lei Yan; Junji Fukuda

STAR Protocols (Jun 2022)

Luciferase assay system to monitor fibroblast growth factor signal disruption in human iPSCs

Seiya Kanno,
Kashu Mizota,
Yusuke Okubo,
Tatsuto Kageyama,
Lei Yan,
Junji Fukuda

Affiliations

Seiya Kanno: Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya Ward, Yokohama, Kanagawa 240-8501, Japan; Division of Cellular & Molecular Toxicology, Center for Biological Safety & Research, National Institute of Health Sciences, 3-25-26 Tono-machi, Kawasaki-ku, Kawasaki, Kanagawa 210-9501, Japan; TechnoPro, Inc., 6-10-1 Roppongi, Minato City, Tokyo 106-6135, Japan
Kashu Mizota: Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya Ward, Yokohama, Kanagawa 240-8501, Japan
Yusuke Okubo: Division of Cellular & Molecular Toxicology, Center for Biological Safety & Research, National Institute of Health Sciences, 3-25-26 Tono-machi, Kawasaki-ku, Kawasaki, Kanagawa 210-9501, Japan; Corresponding author
Tatsuto Kageyama: Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya Ward, Yokohama, Kanagawa 240-8501, Japan; Kanagawa Institute of Industrial Science and Technology (KISTEC), 3-2-1 Sakado, Takatsu Ward, Kawasaki, Kanagawa 213-0012, Japan
Lei Yan: Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya Ward, Yokohama, Kanagawa 240-8501, Japan
Junji Fukuda: Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya Ward, Yokohama, Kanagawa 240-8501, Japan; Kanagawa Institute of Industrial Science and Technology (KISTEC), 3-2-1 Sakado, Takatsu Ward, Kawasaki, Kanagawa 213-0012, Japan; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101439

Abstract

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Summary: We describe a protocol for a live-cell luciferase assay system for continuously monitoring fibroblast growth factor (FGF) signal disruption in human-induced pluripotent stem cells (iPSCs). Signal disrupting effects of chemicals are used as an indicator to evaluate toxicity. The assay is reliably predictive of the effects of limb malformation chemicals (AUC = 0.93). The current approach is limited to FGF signal disruption, and combinations with other types of signaling will be required to detect the effects of different toxicants.For complete details on the use and execution of this protocol, please refer to Kanno et al. (2022a). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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