Abstract: Objective To investigate lipopolysaccharide (LPS)-induced changes of cytoskeletal filamentous actin in primary
isolated pulmonary microvascular endothelial cells (PMVECs) from wild-type and RAGE knock-out mouse. Methods The
lungs of wild-type and RAGE knock-out mice were digested with collagenase type I to obtain endothelial cells purified by
anti-CD31-coupled magnetic beads. The PMVEC identified by factor VIII labeling were stimulated with LPS at different
concentrations and the changes of filamentous actin were observed by confocal microscopy. Results The cultured primary cells
showed typical endothelial cell phenotype as examined with factor VIII labeling. LPS stimulation caused rearrangement of the
cytoskeletal filament F-actin in wild-type mouse PMVECs with stress fiber formation, but such changes were not obvious in
RAGE knock-out mouse PMVECs. Conclusion Mouse PMVECs of a high purity can be obtained by immune magnetic beads.
RAGE is involved in LPS-induced destruction of mouse PMVEC cytoskeletons.