The Botulinum Journal, 2011 Vol.2 No.1, pp.30 - 48
Addresses: Division of Bacteriology, National Institute for Biological Standards and Control, Health Protection Agency, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. ‘ MRC Centre for Developmental Neurobiology, King’s College London, New Hunt’s House, Guy’s Campus, London SE1 1UL, UK. ‘ King’s College London, The Graduate School, Waterloo Bridge, 150 Stamford Street, London, SE1 9NN, UK. ‘ Cell Biology Imaging, National Institute for Biological Standards and Control, Health Protection Agency, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. ‘ Division of Bacteriology, National Institute for Biological Standards and Control, Health Protection Agency, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK
Abstract: Research on more sensitive cell based assays is essential to assess neutralising activities of Botulinum neurotoxins antibodies. A reproducible and robust differentiation protocol was applied on human neuroblastoma SH-SY5Y cells generating neuronal cultures stable for 2–3 weeks. The presence of specific neuronal markers was assessed with labelled antibodies. Accurate and reproducible noradrenaline release levels were obtained which were significantly higher in differentiated cells and correlated with vesicle turnover assessments. The suitability of differentiated SH-SY5Y cells for detecting low levels of Botulinum type A and for toxin neutralisation assay was evidenced using functional end points relevant to Botulinum neurotoxicity.
Keywords: BoNT; botulinum neurotoxins; neurotransmitter release; SH-SY5Y cell line; human neuroblastoma cells; differentiation; cell based assay; synaptic vesicles recycling; neuronal cells.
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