Background

Studying 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 Method

We 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.

Results

Our 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 Methods

In comparison to existing techniques to model neuronal ageing our method is cost effective, requiring no specialist equipment or growth factors.

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.

中文翻译：

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在没有神经营养因子的情况下长期培养 SH-SY5Y 神经母细胞瘤细胞：一种新的神经元衰老模型

背景

由于全球人口老龄化，研究人类老龄化变得越来越重要。然而，它面临着产生老化表型的冗长实验的挑战。通常，重现衰老的标志需要复杂的经验条件，这可能会混淆数据解释。事实上，许多研究使用寿命相对较短的整个生物体，这与人类衰老的相关性可能很小或有限。已经广泛使用细胞系来研究人类体细胞的衰老，但人类神经元衰老的建模在体外有些复杂。

新方法

我们培养了充分表征的 SH-SY5Y 人类神经细胞系，以产生分化为表达神经元表型的细胞的高纯度培养物，并设计了一个方案来维持这些细胞的培养，直到它们积累细胞衰老的生物标志物。

结果

我们的数据验证了一种新颖而简单的技术，可有效分化和长期维持 SH-SY5Y 细胞，表达神经元分化的标志物并证明培养物中的电活动。随着体外时间的推移，这些细胞逐渐积累衰老标志物，例如活性氧的产生增加和氧化损伤的积累。

与现有方法的比较

与现有的神经元老化建模技术相比，我们的方法具有成本效益，不需要专业设备或生长因子。

结论

我们证明了在这些培养条件下生长的 SH-SY5Y 细胞代表了一种简单的神经元衰老模型，适用于细胞生物学、生化和电生理学研究。