Embryonic Stem Cell-Derived Neurons Grown on Multi-Electrode Arrays as a Novel In vitro Bioassay for the Detection of Clostridium botulinum Neurotoxins

Jenkinson, Stephen P.; Grandgirard, Denis; Heidemann, Martina; Tscherter, Anne; Avondet, Marc-André; Leib, Stephen (2017). Embryonic Stem Cell-Derived Neurons Grown on Multi-Electrode Arrays as a Novel In vitro Bioassay for the Detection of Clostridium botulinum Neurotoxins. Frontiers in Pharmacology, 8(73) Frontiers 10.3389/fphar.2017.00073

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Clostridium botulinum neurotoxins (BoNTs) are the most poisonous naturally occurring protein toxins known to mankind and are the causative agents of the severe and potentially life-threatening disease botulism. They are also known for their application as cosmetics and as unique bio-pharmaceuticals to treat an increasing number of neurological and non-neurological disorders. Currently, the potency of biologically active BoNT for therapeutic use is mainly monitored by the murine LD50-assay, an ethically disputable test causing suffering and death of a considerable number of mice. The aim of this study was to establish an in-vitro assay as an alternative to the widely used in-vivo mouse bioassay. We report a novel BoNT detection assay using mouse embryonic stem cell-derived neurons (mESN) cultured on multi-electrode arrays. After 21 days in culture, the mESN formed a neuronal network showing spontaneous bursting activity based on functional synapses and express the necessary target proteins for BoNTs. Treating cultures for 6 h with 16.6 pM of BoNT serotype A and incubation with 1.66 pM BoNT/A or 33 Units/ml of Botox® for 24 h lead to a significant reduction of both spontaneous network bursts and average spike rate. This data suggests that mESN cultured on multi-electrode arrays pose a novel, biologically relevant model that can be used to detect and quantify functional BoNT effects, thus accelerating BoNT research while decreasing animal use.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Research
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Physiology
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Grandgirard, Denis; Heidemann, Martina; Tscherter, Anne and Leib, Stephen

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology

ISSN:

1663-9812

Publisher:

Frontiers

Funders:

[4] Swiss National Science Foundation
[UNSPECIFIED] Swiss Federal Office for Civil Protection

Language:

English

Submitter:

Stephen Leib

Date Deposited:

09 Oct 2017 17:56

Last Modified:

09 Oct 2017 18:02

Publisher DOI:

10.3389/fphar.2017.00073

BORIS DOI:

10.7892/boris.96480

URI:

https://boris.unibe.ch/id/eprint/96480

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