Abstract
BackgroundStudying human ageing is of increasing importance due to the worldwide ageing population. However, it faces the challenge of lengthy experiments to produce an ageing phenotype. Often, to recreate the hallmarks of ageing requires complex empirical conditions that can confound data interpretation. Indeed, many studies use whole organisms with relatively short life spans, which may have little, or limited, relevance to human ageing. There has been extensive use of cell lines to study ageing in human somatic cells, but the modelling of human neuronal ageing is somewhat more complex in vitro. New MethodWe cultured the well-characterised SH-SY5Y human neural cell line to produce high purity cultures of cells differentiated to express a neuronal phenotype, and designed a protocol to maintain these cells in culture until they accumulated biomarkers of cellular ageing. ResultsOur data validate a novel and simple technique for the efficient differentiation and long-term maintenance of SH-SY5Y cells, expressing markers of neuronal differentiation and demonstrating electrical activity in culture. Over time in vitro, these cells progressively accumulate markers of ageing such as enhanced production of reactive oxygen species and accumulation of oxidative damage. Comparison to Existing MethodsIn comparison to existing techniques to model neuronal ageing our method is cost effective, requiring no specialist equipment or growth factors. ConclusionsWe demonstrate that SH-SY5Y cells, grown under these culture conditions, represent a simple model of neuronal ageing that is amenable to cell biological, biochemical and electrophysiological investigation.
Highlights
Human neuroblastoma cell lines, such as SH-SY5Y, are a widely used in vitro model system, and have been utilised for a plethora of neuro science applications including cell viability assays (Jaworska-Feil et al, 2010; Funakohi-Tago et al, 2018), neuronal ultrastructure (Cheng et al, 2020), studies of neurophysiology (Tosetti et al, 1998; Oz and Çelik, 2016) and testing of pharmacological preparations (Cheung et al, 2009; Jiang et al, 2018)
Two weeks after retinoic acid withdrawal there were clear differences in the number of cells surviving in the cultures (Fig. 1A) with no cells surviving in serum-free cultures
Levels of cell death are not increasing with time in vitro implying that the cell popu lation has stabilised by this time in culture
Summary
Human neuroblastoma cell lines, such as SH-SY5Y, are a widely used in vitro model system, and have been utilised for a plethora of neuro science applications including cell viability assays (Jaworska-Feil et al, 2010; Funakohi-Tago et al, 2018), neuronal ultrastructure (Cheng et al, 2020), studies of neurophysiology (Tosetti et al, 1998; Oz and Çelik, 2016) and testing of pharmacological preparations (Cheung et al, 2009; Jiang et al, 2018). To overcome the issues of having a heterogeneous cell population, these cells can be induced, by a variety of protocols, to differentiate into a mature neuron-like phenotype This is characterised by exit from the cell cycle, the formation of a rounded cell body coupled with extension of neurites and expression of neuron-specific markers such as βIII-tubulin, NeuN, neurofilament and synaptic vesicle marker Sv2 (Agholme et al, 2010). Conclusions: We demonstrate that SH-SY5Y cells, grown under these culture conditions, represent a simple model of neuronal ageing that is amenable to cell biological, biochemical and electrophysiological investigation
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
