Oral or Poster Presentation
Concurrent Session 3B - Maternal Fetal Medicine
Introduction: Over 80% of perinatal brain injuries lead to neurodevelopmental disabilities. Therefore, preventive approaches are needed. One major factor is placental insufficiency which results in hypoxic-ischemic environment in-utero. Sulforaphane (SFA), derived from cruciferous vegetables, is a phase-II enzyme inducer that enhances anti-oxidant enzymes production. The aim of this study is to explore how SFA affects different brain cell types and the dosing range of SFA for protection/toxicity in normal and oxygen/glucose deprived (OGD) cell cultures.
Methods: We used primary cortical neuronal, astrocyte, and combined cell cultures (co-cultures) from newborn rodents. Cell culture purity was evaluated by Western blot (WB) and immunofluorescence (IF). Cultures were exposed to OGD until 50% cell death (LD50) was achieved. We exposed cultures at LD50 to varying doses of SFA. Controls were not exposed to OGD. Cell viability was assessed by a Live/Dead assay using IF/high-content microscopy and cytotoxicity by Alamar blue. One Way ANOVA and Dunette's Multiple Comparison were used for statistics.
Results: Primary cortical neuronal, astroglial, and co-cultures have been established. Using WB and IF for cell specific markers, we show: neuronal cultures with minimal contamination of astrocytes and microglia; astrocyte cultures with minimal contamination of neurons and microglia. The co-cultures showed markers for all cell types. We determined the LD50 to be 2 hours for neurons, 4 hours for astrocytes, and 10 hours for co-cultures. At LD50, SFA was protective at 2.5uM for neurons (p<0.01), astrocytes, and co-cultures. Significant toxicity of SFA in control cultures was seen at doses ≥100uM (p<0.01) for neurons, and ≥50uM (p<0.01) for both astrocytes and co-cultures.
Conclusion: These findings suggest that SFA shows promise as a preventative agent for fetal ischemic brain injury and that dosing parameters are required for safety. Future studies will determine the safety/efficacy of SFA in a rat animal model of placental insufficiency.