• Optimizing Isolation and Culture of Primary Microglia

      Doughty, Deanna; Department of Biological Sciences; American Cancer Society -- Institutional Research Grant; Augusta University PILOT Award (Augusta University, 2018-12)
      Glioblastoma (GBM) is the most aggressive and common adult brain tumor subtype, with the majority of patients surviving less than one year. The GBM microenvironment is composed of tumor cells as well as non-cancerous cells, such as microglia, a component of the immune system in the brain. To better understand the role of microglia in GBM, we have optimized in vitroculture conditions for primary microglia. Growing microglia in culture is challenging, but this technique is needed for planned future experiments. Microglia were isolated from mouse neuronal tissue by magnetic bead antibody cell separation using the cellular marker CX3CR1. Isolated microglia were then cultured in various culture conditions, and cellular morphology by light microscopy was used to determine cell health, viability, and activation status. It was determined that the primary microglia grow best in neurobasal media in wells coated with poly-D lysine. Future studies aim to isolate a larger number of cells to allow forco-culture of the inactivated microglia with GBM cells. These results will allow us to better understand the role that microglia play in GBM progression.