Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078,China

Objective To establish a method for the isolation and culture of mouse liver sinusoidal endothelial cells (LSEC), and to study their biological characteristics.MethodsLiver tissue from mouse was treated with a solution containing 2 U/mL dispase, and then the cell suspension was separated by Percoll density gradient centrifugation. LSEC was purified by culturing in an endothelial cell selective medium and passaged with enzyme discrepancy digestion. LSEC immunophenotypic characteristics were analyzed by using cytoimmunostaining. The ultrastructures of LSEC, the ability of uptaking the acetylated lowdensity lipoprotein (acetylated LDL), and the vascular structures in vitro were detected.ResultsThe isolated cells showed the typical cobblestone morphology characteristic of endothelial cells. These cells expressed factor Ⅷ but not CD31, CD90, and CK19. They were able to uptake the acetylated lowdensity lipoprotein and to form vascular structure. The results of electron microscope showed that these cells possessed typical transcellular fenestrations with a sinusoidal origin.Conclusion We develop a method for the isolation and culture of LSEC with typical morphologic and phenotypic characteristics.