A highly specific isolation of rat sinusoidal endothelial cells by the immunomagnetic bead method using SE-1 monoclonal antibody |
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Authors: | Tokairin Takuo Nishikawa Yuji Doi Yuko Watanabe Hitoshi Yoshioka Toshiaki Su Mu Omori Yasufumi Enomoto Katsuhiko |
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Affiliation: | Department of Pathology, Akita University School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan. |
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Abstract: | BACKGROUND/AIMS: To develop a specific isolation method of hepatic sinusoidal endothelial cells (SEC), we applied the immunomagnetic method using a monoclonal antibody (SE-1) that recognizes a membranous antigen expressed only in rat SEC.METHODS: Cells were isolated by incubating mixed non-parenchymal cells, which were obtained by collagenase digestion of the liver, with SE-1-conjugated superparamagnetic polystyrene beads. The conventional Percoll method was also performed in parallel to compare with the immunomagnetic method. The isolated cells were cultured on glass coverslips coated with type I collagen in the presence of various growth factors for 6 days.RESULTS: Approximately 98% of the isolated cells were positive for SE-1 and the contamination of Kupffer cells or stellate cells was less than 1%. The purity was significantly better than that obtained by the Percoll method. The cultured cells showed typical SEC features, such as sieve plates and uptake of acetylated low-density lipoprotein. Although the cells continuously underwent apoptotic cell death after 2 days, they started robust cell growth after 3 days and were well maintained during the culture period.CONCLUSIONS: Our simple and specific isolation method enables us to culture SEC with high purity and should be useful for the biological analysis of SEC. |
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Keywords: | Sinusoidal endothelial cell Immunomagnetic bead isolation SE-1 antibody Tissue culture Apoptosis Cell growth |
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