Analysis of motor neurons differentiated from human induced pluripotent stem cells for the use in cell-based Botulinum neurotoxin activity assays
| Autor: | Schenke, Maren, Schjeide, Brit-Maren, Püschel, Gerhard Paul, Seeger, Bettina |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
potency assessment
Botulinum Toxins Dose-Response Relationship Drug induced pluripotent stem cells Neurogenesis lcsh:R Neurotoxins Acetylcholine Release Inhibitors lcsh:Medicine Animal Testing Alternatives cell-based in vitro assay Article Cell Line Neural Stem Cells Botulinum neurotoxin Humans motor neurons Biological Assay Institut für Ernährungswissenschaft ddc:610 610 Medizin und Gesundheit Institut für Biochemie und Biologie |
| Zdroj: | Toxins Volume 12 Issue 5 Toxins, Vol 12, Iss 276, p 276 (2020) |
| Popis: | Botulinum neurotoxins (BoNTs) are potent neurotoxins produced by bacteria, which inhibit neurotransmitter release, specifically in their physiological target known as motor neurons (MNs). For the potency assessment of BoNTs produced for treatment in traditional and aesthetic medicine, the mouse lethality assay is still used by the majority of manufacturers, which is ethically questionable in terms of the 3Rs principle. In this study, MNs were differentiated from human induced pluripotent stem cells based on three published protocols. The resulting cell populations were analyzed for their MN yield and their suitability for the potency assessment of BoNTs. MNs produce specific gangliosides and synaptic proteins, which are bound by BoNTs in order to be taken up by receptor-mediated endocytosis, which is followed by cleavage of specific soluble N-ethylmaleimide-sensitive-factor attachment receptor (SNARE) proteins required for neurotransmitter release. The presence of receptors and substrates for all BoNT serotypes was demonstrated in MNs generated in vitro. In particular, the MN differentiation protocol based on Du et al. yielded high numbers of MNs in a short amount of time with high expression of BoNT receptors and targets. The resulting cells are more sensitive to BoNT/A1 than the commonly used neuroblastoma cell line SiMa. MNs are, therefore, an ideal tool for being combined with already established detection methods. Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe; 1083 |
| Databáze: | OpenAIRE |
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