Acquiring Kupffer cells in mice using a MACS-based method.

Authors :: Zhang C
Lu Y
Zhou H
Lu H
Qian X
Liu X
Wang X
Ding Z
Zhang F
Lu L
Source :: Transplantation proceedings [Transplant Proc] 2015 Mar; Vol. 47 (2), pp. 553-7.
Publication Year :: 2015
Abstract: Objective: This study sought to establish a new method to isolate Kupffer cells (KCs) by magnetic activated cell sorting (MACS).<br />Methods: Nonparenchymal 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.<br />Results: Compared 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.<br />Conclusions: With 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.<br /> (Copyright © 2015 Elsevier Inc. All rights reserved.)