关键词: 细胞培养, 兔血管内皮细胞, 生物学特性

Abstract: The aim of the study was to develop a method for the isolation, identification and primary culture of rabbit vascular endothelial cells(RVECs) as a modle for studying the pathologenicity of rabbit haemorrheagic disease virus on RVECs.RVECs were dissociated by trypsin digestion and tissue pieces culture method and cultured in vitro.The REVCs were identified by factor Ⅷ related antigen immunohistochemical staining and CD34 indirect immunofluorescence assay.The result showed that the cells which gained by digesting method adhered about 20 h, and filled the basis of petri dish after 1w with high activity.The cells which isolated by tissue pieces culture method grew from original about 2 weeks and filled the basis of petri dish 3 weeks later, and these cells were contaminated with smooth muscle cells and fibroblast cells, while they had higher vitality.After cells were passaged and cultured, they exhibited as polygon and spindle-shaped cells and displayed cobblestone-likemorphology after fusion to monolayer.Immunohistochemical staining and indirect immunofluorescence revealed that the expression of factor Ⅷ and CD34 was positive.

Key words: cell culture, rabbit vascular endothelial cells, biological characteristics