Acquiring Kupffer Cells in Mice Using a MACS-Based Method

• With the 2-step method of immunomagnetic beads, we provide a new method by which Kupffer cells can be obtained from mouse liver with high purity and distinct phenotype of F4/80+ CD11c−.

ObjectiveThis study sought to establish a new method to isolate Kupffer cells (KCs) by magnetic activated cell sorting (MACS).MethodsNonparenchymal cells were acquired from C57BL/6 mice livers by a perfusion system in vivo and then stained with F4/80+ fluorescein isothiocyanate and CD11c− phycoerythrin antibodies. After incubating with immunomagnetic beads, F4/80+CD11c− KCs were obtained by MACS selection. The purity was evaluated by flow cytometry, and the morphological features and vitality were analyzed in in vitro cultures.ResultsCompared with traditional methods, acquiring KCs by MACS was characterized by economy, efficiency, and high purity. The F4/80+CD11c− KCs cultured in vitro also showed the typical adherent shape and excellent phagocytic ability.ConclusionsWith the 2-step method using immunomagnetic beads, we provide a new method by which KCs can be obtained from mouse liver with high purity and distinct phenotype of F4/80+ CD.