A highly specific isolation of rat sinusoidal endothelial cells by the immunomagnetic bead method using SE-1 monoclonal antibody.
OBJECTIVE
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.
Tokairin
Takuo
T
Department of Pathology, Akita University School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan.
Nishikawa
Yuji
Y
Doi
Yuko
Y
Watanabe
Hitoshi
H
Yoshioka
Toshiaki
T
Su
Mu
M
Omori
Yasufumi
Y
Enomoto
Katsuhiko
K
D000818:Animals; D000911:Antibodies, Monoclonal; D017209:Apoptosis; D002455:Cell Division; D002478:Cells, Cultured; D004727:Endothelium / Q000166:cytology; D018189:Immunomagnetic Separation; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology* / Q000276:immunology*; D008297:Male; D010641:Phenotype; D011205:Povidone; D051381:Rats; D011916:Rats, Inbred F344; D012822:Silicon Dioxide
D000911:Antibodies, Monoclonal; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone