Isolation and Characterization of Primary Microglial Cells From Mouse Cortical Tissue

The neuronal, actin cytoskeleton regulates numerous processes, including cell shape modulation, migration, and maintenance of polarity, that aid in regulating homeostasis of the cell. The cytoskeleton is especially important for cell motility, morphology, and immune responses. A regulator of actin cytoskeleton is cofilin, an actin-binding protein that disassembles the actin filaments. Several studies have shown that rapid cycles of actin assembly and disassembly depend on cofilin because it stimulates disassembly. This can possibly explain why cofilin is found in nearly all eukaryotes and why it is essential for cell viability. In previous semesters, our lab has shown that HIV-1 infection resulted in a pro-inflammatory environment that led to modulations in phagocytic activity in the BV-2 microglial cell line. Our aim is to isolate primary macrophages from mouse cortical tissue because it acts as a better model to compare the physiological changes seen in humans. Our novel technique will provide an insight into possible contributory factors of HIV associated neurocognitive diseases and novel pathways for therapeutic targets. In order to do this, we use immunofluorescence staining of slides to characterize cell type and activatio