title: Human Induced Pluripotent Stem Cell Derived Neuronal Cells Cultured on Chemically-Defined Hydrogels for Sensitive In Vitro Detection of Botulinum Neurotoxin.
abstract: Botulinum neurotoxin (BoNT) detection provides a useful model for validating cell-based neurotoxicity screening approaches, as sensitivity is dependent on functionally competent neurons and clear quantitative endpoints are available for correlating results to approved animal testing protocols. Here, human induced pluripotent stem cell (iPSC)-derived neuronal cells were cultured on chemically-defined poly(ethylene glycol) (PEG) hydrogels formed by "thiol-ene" photopolymerization and tested as a cell-based neurotoxicity assay by determining sensitivity to active BoNT/A1. BoNT/A1 sensitivity was comparable to the approved in vivo mouse bioassay for human iPSC-derived neurons and neural stem cells (iPSC-NSCs) cultured on PEG hydrogels or treated tissue culture polystyrene (TCP) surfaces. However, maximum sensitivity for BoNT detection was achieved two weeks earlier for iPSC-NSCs that were differentiated and matured on PEG hydrogels compared to TCP. Therefore, chemically-defined synthetic hydrogels offer benefits over standard platforms when optimizing culture conditions for cell-based screening and achieve sensitivities comparable to an approved animal testing protocol.
authors.affiliation: Department of Bacteriology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: Sabine
authors.initials: S
authors.lastname: Pellett
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Schwartz
authors.affiliation: Department of Bacteriology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: William H
authors.initials: WH
authors.lastname: Tepp
authors.affiliation: MTI-GlobalStem, Inc. Gaithersburg, Maryland, United States of America.
authors.forename: Richard
authors.initials: R
authors.lastname: Josephson
authors.affiliation: Department of Bacteriology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: Jacob M
authors.initials: JM
authors.lastname: Scherf
authors.affiliation: Department of Bacteriology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: Christina L
authors.initials: CL
authors.lastname: Pier
authors.affiliation: Department of Cell and Regenerative Biology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.|Morgridge Institute for Research, Madison, Wisconsin, United States of America.|Department of Molecular, Cellular, and Developmental Biology, University of California at Santa Barbara, Santa Barbara, California, United States of America.
authors.forename: James A
authors.initials: JA
authors.lastname: Thomson
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.|Department of Orthopedics and Rehabilitation, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: William L
authors.initials: WL
authors.lastname: Murphy
authors.affiliation: Department of Bacteriology, University of Wisconsin at Madison, Madison, Wisconsin, United States of America.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Johnson
mesh_terms: D000818:Animals; D001905:Botulinum Toxins / Q000494:pharmacology*; D019274:Botulinum Toxins, Type A / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects*; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000187:drug effects*; D011092:Polyethylene Glycols / Q000737:chemistry
chemical_list: D020100:Hydrogels; D011092:Polyethylene Glycols; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A