Innate immune cells are notoriously difficult to transfect; however, retroviruses can be used to stably integrate genes of interest into the host genome of primary or immortalized immune cells resulting in the generation of reporter cells. Here, we provide a detailed protocol covering the production of retroviruses, retroviral infection of innate immune target cells (including isolation and differentiation of murine bone marrow cells to macrophages), and several methods for enrichment of positively transduced cells.
Keywords: Bone marrow isolation; Bone marrow-derived macrophages; Fluorescent protein; Infection; Innate immune reporter cells; Innate immunity; Retrovirus; Transduction.