Single cell preparations of Mycobacterium tuberculosis damage the mycobacterial envelope and disrupt macrophage interactions

In order to investigate the impact of using in vitro techniques to generate single cell suspensions of Mycobacterium tuberculosis (Mtb) on macrophage gene expression, we compared uninfected bone marrow derived macrophages to macrophages infected with Mtb that was prepared using gentle sonication followed by low-speed centrifugation (so/sp) or passage through a 5 µm syringe filter (5µmF). Overall design: Comparative gene expression profiling analysis of RNA-seq data for wild-type murine macrophages were uninfected or infected with Mtb H37Rv at a multiplicity of infection of 5 followed by RNA sequencing at 72 hours post infection. 5 experimental replicates were performed for each group.