C Zhang1, Y Lu1, H Zhou1, H Lu1, X Qian1, X Liu1, X Wang1, Z Ding1, F Zhang1, L Lu2. 1. Translational Medicine Research Center of Jiangning Hospital and Liver Transplantation Center of First Affiliated Hospital, Nanjing Medical University, Nanjing, China. 2. Translational Medicine Research Center of Jiangning Hospital and Liver Transplantation Center of First Affiliated Hospital, Nanjing Medical University, Nanjing, China. Electronic address: lvling@njmu.edu.cn.
Abstract
OBJECTIVE: This study sought to establish a new method to isolate Kupffer cells (KCs) by magnetic activated cell sorting (MACS). 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. 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. 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.
OBJECTIVE: This study sought to establish a new method to isolate Kupffer cells (KCs) by magnetic activated cell sorting (MACS). 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. 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. 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.
Authors: Richard J P Smith; Alessandro Faroni; James R Barrow; Jamie Soul; Adam J Reid Journal: Stem Cell Res Ther Date: 2021-03-02 Impact factor: 6.832