Extracellular vesicles from water kefir can interact with human neurons in vitro: a potential explanation for the role of probiotics consumption in mental health

Lora J. Kasselman; Saba Ahmed; Ariel De Leon; Maryann Johnson; Ankita Srivastava; Apoorva Vashisht; Heather A. Renna; Thomas Palaia; Aaron Pinkhasov; Allison B. Reiss; Lora J. Kasselman; Saba Ahmed; Ariel De Leon; Maryann Johnson; Ankita Srivastava; Apoorva Vashisht; Heather A. Renna; Thomas Palaia; Aaron Pinkhasov; Allison B. Reiss

doi:10.3934/Neuroscience.2025019

AIMS Neuroscience

2025, Volume 12, Issue 3: 351-368. doi: 10.3934/Neuroscience.2025019

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Research article

Extracellular vesicles from water kefir can interact with human neurons in vitro: a potential explanation for the role of probiotics consumption in mental health

1.
Hackensack Meridian Jersey Shore University Medical Center, Hackensack, NJ, USA
2.
Department of Foundations of Medicine, NYU Grossman Long Island School of Medicine, Mineola, NY, 11501, USA
3.
Department of Medicine, NYU Grossman Long Island School of Medicine, Mineola, NY, 11501, USA
4.
Department of Psychiatry, NYU Grossman Long Island School of Medicine, Mineola, NY, 11501, USA

Received: 07 April 2025 Revised: 04 August 2025 Accepted: 11 August 2025 Published: 21 August 2025

Major depressive disorder is one of the most burdensome mental health disorders. Probiotics have been shown to ameliorate depressive symptoms, though the mechanism remains unclear. This study was conducted to investigate whether extracellular vesicles (EVs) extracted from the probiotic beverage water kefir could influence gene and protein expression in human-derived neuroblastoma cells in vitro. EVs were extracted from lab-cultured water kefir and a control solution without water kefir grains by ultracentrifugation. Water kefir vesicles were imaged via electron microscopy. Neuroblastoma, microglia, and neuroblastoma-microglia co-cultures were exposed to water kefir EVs or negative control medium. Uptake of water kefir EVs was identified by microscopy. All conditions were quantified for brain derived neurotrophic factor, fractalkine, and synaptophysin RNA and protein. Data were analyzed using factorial ANOVAs with significance set at 0.05. Water kefir vesicles were taken up by neuroblastoma cells, and incubation in neuroblastoma-microglia co-culture resulted in significantly higher levels of fractalkine protein compared to media-only control (p = 0.029). To our knowledge, this is the first study to identify potential interactions between EVs derived from the probiotic beverage water kefir and human neuronal cells. Further research is needed to fully elucidate the role played by probiotic-derived EVs in human health.
- mental health,
- depression,
- fermentation,
- culture,
- cognition,
- gut-brain axis
Citation: Lora J. Kasselman, Saba Ahmed, Ariel De Leon, Maryann Johnson, Ankita Srivastava, Apoorva Vashisht, Heather A. Renna, Thomas Palaia, Aaron Pinkhasov, Allison B. Reiss. Extracellular vesicles from water kefir can interact with human neurons in vitro: a potential explanation for the role of probiotics consumption in mental health[J]. AIMS Neuroscience, 2025, 12(3): 351-368. doi: 10.3934/Neuroscience.2025019

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Abstract

Major depressive disorder is one of the most burdensome mental health disorders. Probiotics have been shown to ameliorate depressive symptoms, though the mechanism remains unclear. This study was conducted to investigate whether extracellular vesicles (EVs) extracted from the probiotic beverage water kefir could influence gene and protein expression in human-derived neuroblastoma cells in vitro. EVs were extracted from lab-cultured water kefir and a control solution without water kefir grains by ultracentrifugation. Water kefir vesicles were imaged via electron microscopy. Neuroblastoma, microglia, and neuroblastoma-microglia co-cultures were exposed to water kefir EVs or negative control medium. Uptake of water kefir EVs was identified by microscopy. All conditions were quantified for brain derived neurotrophic factor, fractalkine, and synaptophysin RNA and protein. Data were analyzed using factorial ANOVAs with significance set at 0.05. Water kefir vesicles were taken up by neuroblastoma cells, and incubation in neuroblastoma-microglia co-culture resulted in significantly higher levels of fractalkine protein compared to media-only control (p = 0.029). To our knowledge, this is the first study to identify potential interactions between EVs derived from the probiotic beverage water kefir and human neuronal cells. Further research is needed to fully elucidate the role played by probiotic-derived EVs in human health.

Acknowledgments

This work was supported by The Alzheimer's Foundation of America (AWD00004772) and The Herb and Evelyn Abrams Family Amyloid Research Fund. With special thanks to Edmonds Bafford, Dolores McCormack and Deborah Zimmer.

Conflict of interest

The authors declare no conflict of interest.

Authors' contributions

Lora J. Kasselman: Conceptualization, design, validation, formal analysis, writing – original draft, writing – review and editing; Saba Ahmed: Investigation, methodology, validation, formal analysis; Ariel De Leon: Writing – original draft, writing – review and editing, software, data curation; Maryann Johnson: Investigation, methodology; Ankita Srivastava: Formal analysis, methodology, referencing; Apoorva Vashisht: Investigation, methodology, writing – review and editing; Heather A. Renna: Investigation, methodology; Thomas Palaia: Electron microscopy, investigation, methodology; Aaron Pinkhasov: Conceptualization, writing- review and editing; Allison B. Reiss: Conceptualization, project administration, design, supervision, writing – original draft, writing – review and editing, funding acquisition. All authors read and approved the final version of the manuscript.

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