Objective     To establish a comprehensive characterization protocol of rat pulmonary microvascular endothelial cells (RPMVECs) cultured by tissue-sticking method.      Methods     RPMVECss were cultured with peripheral lung tissue-sticking method. Histological sections from peripheral lung tissue pieces used for cell culture were examined. With rat pulmonary artery smooth muscle cells and human umbilical vein endothelial cells as controls

the cultured cells were identified by immunocytochemical staining using CD34 antibody

lectin from Bandeiraea simplicifolia and factor Ⅷ related antigen. The cell morphology and ultrastructure were observed with inverted light microscope and transmission electron microscope respectively.      Results     Histological sections showed that tissue pieces were scissored from peripheral lung lobes accurately. The cultured cells had characterization of binding lectin from Bandeiraea simplicifolia and positive immunocytochemical staining with CD34 antibody

but negative for factor Ⅷ related antigen. Weibel-Palade bodies were not found in cells.      Conclusion     Factor Ⅷ related antigen and Weibel-Palade body are not ideal indexes for RPMVECs identification. The combination of peripheral lung tissue section

CD34 immunocytochemical staining and lectin from Bandeiraea simplicifolia binding assay provides a simple and reasonable comprehensive characterization protocol for RPMVECs.