Abstract T P343: Resveratrol Rescues Neuron Survival in Pimary Postnatal Cultures
Objective: Our goal is to identify supplements that can improve neuronal survival of postnatal neuronal cultures. Here, we hypothesize that resveratrol could rescue neurons that are stressed during the brain dissociation and plating procedures.
Background: The standardized B27 supplement with neurobasal medium has been used to facilitate neuronal growth and attenuates glial proliferation. We have documented that resveratrol (3,4’5 trihydroxystilbene), a natural polyphenol and a bioactive component of red wine, have numerous neuroprotective properties both in vivo and after oxygen-glucose deprivation protocols. We have proposed that this stilbene would not likely act as a direct antioxidant but act as a bioactive agent capable to induce an intracellular cytoprotective cascade.
Methods: Postnatal cultures of cortical neurons were derived from 0- to 1-day-old C57BL/6 mice. Brains were dissociated and neurons plated and maintained in three different media: neurobasal medium without any supplement (NBM), neurobasal medium without any supplement but containing resveratrol 10μM (NBM+R10μM), and neurobasal medium with B27 supplement (NBM+B27). Neuronal survival was assessed with the live/dead cell calcein AM/EthD-1), lactate dehydrogenase (LDH) and (MTT) assays after plating in these three different media.
Results: Significant increase in live cells (52.9+/-5.5% and 53.6+/-4.5%, p<0.0001) was recorded in cultures incubated in NBM+R10μM and NBM+B27 respectively, when compared to NBM by calcein-AM assay and increase in viability (50.8+/-8.5% and 54.3+/-6.5%) by MTT assay. Significant decrease in dead cells (73.9+/-6.3% and 74.7+/-5.3%, p<0.0001) was recorded in cultures incubated in NBM+R10μM and NBM+B27 respectively, when compare to NBM by EthD-1 assay and decrease in cytotoxicity (17.9+/-2.5% and 19.4+/-2.8) by LDH assay.
Conclusions: Results indicated that growth of primary neurons in resveratrol containing medium was most effective suggesting that resveratrol itself is sufficient to enhance neuronal attachment and survival during plating. Efforts are currently being conducted to characterize the mechanism by which resveratrol can limit cell death and increase neuron survival.
Author Disclosures: M. Agrawal: None. S. Dore: None.
- © 2014 by American Heart Association, Inc.