Comparison of medium supplements in terms of the effects on the differentiation of SH-SY5Y human neuroblastoma cell line

Objective: Human SH-SY5Y cell line has been frequently used for in vitro experiments in neuroscience-related research. To reflect a better neuronal characteristic the cell line needs to a differentiation. To compare the results obtained from in vitro models, the similarity of the phenotype and characteristic of the cells has great importance. However, many studies have been performed using the different medium ingredients which affect the differentiation progress of the cells. Therefore, we aimed to compare generally used differentiation mediums, contain only retinoic acid (RA) and supplemented with different ingredients, in the aspect of neuron-like phenotype characteristics and stability. Materials and Methods: The effects of medium ingredients on differentiation levels were evaluated using morphological changing, neurite length, and immunofluorescence detection of neuronal markers such as NFH, β-III Tubulin, and microtubule-associated protein 2 (MAP2). The stability of differentiated cells was followed microscopically at the 7th, 10th, and 14th days by morphological changings and neurite length using Neuron J software. Results: The results revealed that the cells pretreated with RA for 5-day treatment and followed by 5-day treatment with the mix medium and brain-derived neurotrophic factor (BDNF), provided significantly higher neurite length than the other groups (P < 0.001). In this group, the expressions of β-tubulin III, MAP2, and NF-H were also significantly higher than the control group (P < 0.05, P < 0.001, and P < 0.05, respectively) and differentiated cells were stable until the 7th day. Conclusion: The results demonstrated that enriched mediums are necessary for a better differentiation of SH-SY5Y cells. We recommend 10-day treatment period and using of RA, BDNF, dc-AMP, KCI together in SH-SY5Y cell differentiation