Generation of microglia from peripheral blood mononuclear cells (PBMCs)

Microglia are resident brain macrophages that modulate the release of cytokines to mediate neuroinflammation. Conversely, they may release inflammatory mediators that promote protein aggregation and neuronal damage in neurodegenerative diseases. Furthermore, challenges with obtaining mature microglia reprogrammed from inducible pluripotent stem cells (iPSCs) hinder the ability to accurately understand their role in neurodegenerative diseases in vitro. Our objective, therefore, is to derive phenotypically mature and biologically relevant microglia derived from PBMCs. For the methodology, PBMCs were reprogrammed to microglia using IL-34 and GM-CSF for 2 weeks. We were able to successfully obtain morphologically representative microglia. To further verify the phenotype of the obtained microglia, we performed a flow cytometry where cells expressed Iba1, CD11b, and had no CD45, which is reminiscent of microglia in vivo. Ongoing and future studies will assess the effect of HIV and antiretroviral therapy on resident brain myeloid cells such as microglia an macrophages.