Influence of microbially fermented 2´-fucosyllactose on neuronal-like cell activity in an in vitro co-culture system

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Scope 2´-Fucosyllactose (2´-FL), the most abundant oligosaccharide in human milk, plays an important role in numerous biological functions, including improved learning. It is not clear, however, whether 2´-FL or a cleavage product could influence neuronal cell activity. Thus, we investigated the effects of 2´-FL, its monosaccharide fucose (Fuc), and microbial fermented 2´-FL and Fuc on the parameters of neuronal cell activity in an intestinal–neuronal transwell co-culture system in vitro . Methods Native 13 C-labeled 2´-FL and 13 C-Fuc or their metabolites, fermented with Bifidobacterium (B.) longum ssp . infantis and B. breve , which were taken from the lag-, log- and stationary (stat-) growth phases of batch cultures, were applied to the apical compartment of the co-culture system with Caco-2 cells representing the intestinal layer and all- trans -retinoic acid-differentiated SH-SY5Y (SH-SY5Y ATRA ) cells mimicking neuronal-like cells. After 3 h of incubation, the culture medium in the basal compartment was monitored for 13 C enrichment by using elemental analysis isotope-ratio mass spectrometry (EA-IRMS) and effects on cell viability, plasma, and mitochondrial membrane potential. The neurotransmitter activation (BDNF, GABA, choline, and glutamate) of SH-SY5Y ATRA cells was also determined. Furthermore, these effects were also measured by the direct application of 13 C-2´-FL and 13 C-Fuc to SH-SY5Y ATRA cells. Results While no effects on neuronal-like cell activities were observed after intact 2´-FL or Fuc was incubated with SH-SY5Y ATRA cells, supernatants from the stat-growth phase of 2´-FL, fermented by B. longum ssp. infantis alone and together with B. breve , significantly induced BDNF release from SH-SY5Y ATRA cells. No such effects were found for 2´-FL, Fuc, or their fermentation products from B. breve . The BDNF release occurred from an enhanced vesicular release, which was confirmed by the use of the Ca 2+ -channel blocker verapamil. Concomitant with this event, 13 C enrichment was also observed in the basal compartment when supernatants from the stat-growth phase of fermentation by B. longum ssp. infantis alone or together with B. breve were used. Conclusion The results obtained in this study suggest that microbial products of 2´-FL rather than the oligosaccharide itself may influence neuronal cell activities.

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