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. Overall design: 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.