(P27-038-23) The Impact of Membrane Polyunsaturated Fatty Acid Composition on Neuronal Cell Development

Objectives: This study aimed to investigate the impact of individual polyunsaturated fatty acids (PUFAs) on membrane composition, n-6:n-3 ratio, and morphology during neuronal cell differentiation and development in culture.

Methods: To model neuronal development, SH-SY5Y human neuroblastoma cells were cultured with differentiating media (1% fetal bovine serum (FBS), 10µM retinoic acid) for 5 days to differentiate the cells into the neuronal phenotype. The differentiating media was supplemented with 70µM of α-linolenic acid (ALA), arachidonic acid (AA), or eicosapentaenoic acid (EPA) conjugated to BSA or BSA control with 3 replicates per treatment. Phase-contrast microscopy images were taken daily during the differentiation process to document cell morphology as an indicator of development and were used to quantify the neurite number per cell. Membrane fatty acid (FA) composition was analyzed using gas chromatography (GC) following lipid extraction and FA methylation. Cell viability was determined using colorimetric assays. Statistical analysis was conducted using one-way ANOVA with Tukey’s HSD post-hoc analysis.

Results: With ALA and EPA treatments, the mean concentrations of EPA and n-3 docosapentaenoic acid (DPA) significantly increased, while the mean concentration of ALA significantly increased only with ALA treatment (p < 0.05). With AA treatment, the mean concentration of AA significantly increased whereas the mean concentration of DHA significantly decreased (p < 0.05). The total n-3 PUFAs proportion were significantly increased in ALA and EPA treatments, while total n-6 PUFAs proportions was significantly increased with AA treatment (p < 0.05). The n-6:n-3 ratio was significantly higher with AA treatment and significantly lower with ALA and EPA treatments (p < 0.05). The AA treatment decreased cell viability, while ALA and EPA treatments had no effect. For all treatments, the number of neurites per cell increased from day 1 to day 5 with no FA treatment effects observed.

Conclusions: Media supplementation of ALA, AA, and EPA individually altered the PUFA composition and n-6:n-3 ratio of cultured SH-SY5Y neuronal cell membranes, however FA treatments did not affect the rate of cell development. Further experiments are required to assess functional outcomes and other morphological measures of developing neurons.

Funding Sources: Purdue University