RNA-seq of SCD2-deficient murine bone marrow-derived macrophages

We evaluated the effect of Scd2 ablation on the transcriptome of bone marrow-derived macrophages prepared from mice. We obtained mice with floxed Scd2 alleles from Hide Tsukamoto (Keck School of Medicine from the University of Southern California). We crossed these mice with mice carrying the lysozyme M-Cre transgene to generate mice with myelomonocyte-specific deletion of Scd2. We prepared macrophages from bone marrow. To isolate bone marrow from mice, we sacrificed mice using CO2 euthanasia, dissected out both tibias and femurs, and passed 8 ml of ice-cold DMEM through each bone's medullary cavity. We filtered the bone marrow aspirates through a 40 micrometer nylon mesh strainer, pelleted the cells, and re-suspended the pellet in macrophage differentiation media consisting of DMEM with 10% FBS, 10% CMG-conditioned media, 1% P/S, and 1% L-glut. We replenished differentiation media 3 days after initial plating. After differentiation for 5-6 days, macrophages were maintained in mature macrophage media consisting of DMEM with 10% FBS, 2% CMG-conditioned media, 1% P/S, and 1% L-glut.