miRNA microarray expression from glioblastoma & astrocytes under 100V/m 48hr dcEF stimulation & sham

Direct current electric field mimicking a physiological electrical field from transepithelial potential difference can direct cell migration (electrotaxis)and cellular signaling. While the transcriptome in dcEF has been reported and more studied, the miRNA expression of cells under dcEF stimulation is less understood. We use a reversibly sealed dcEF stimulation bioreactor to apply uniform dcEF to glioblastoma cell lines and primary astrocytes and investigate if dcEF can induce differential expression of cellular miRNA and exosomal miRNA expression that can regulate the gene expression Cells were cultured in the the bioreactor prior to assembly in a physioxia (5%CO2, 5%O2) environment. The bioreactor was assembly submerged in 1X PBS to avoid bubble entrapment and then 100V/m dcEF was applied for 48hours. The conditioned medium containing the exosome was collected and isolated for exosomal RNA. Cellular total RNA was also extracted. Duplicated DNA are tagged and pooled. Then 130ng of RNA of cellular RNA and exosomal RNA were hybridized in miRNA 4.0 microarray. RMA+DABG with normalization and human only miRNA analysis was performed.