中国病理生理杂志2012，Vol.28Issue(1)：186-188,192,4.DOI:10.3969/j.issn.1000-4718.2012.01.037
小鼠肺微血管内皮细胞的培养鉴定及其血管形成功能的研究
Primary culture, identification and in vitro angiogenesis of mouse pulmonary microvascular endothelial cells
摘要
Abstract
AIM:To establish a simple and reliable method for primary culture of mouse pulmonary microvas-cular endothelial cells ( PMVECs ), and to investigate the ability of angiogenesis of the cells in vitro. METHODS:PM-VECs from C57BL/6 new born mice were cultured with peripheral pulmonary tissue pasted method. The cells were observed under inverted phase - contrast microscope and identified by immunocytochemical and immunofluo re scent methods. PMVECs were seeded at the density of 20 000 cells per well in 96 - well plate on top of growth factor - reduced matrigel and incubated for 24 h at 37℃ in 5% CO2. The plates were examined every 2 h for tube formation under an inverted microscope. RESULTS: The primary PMVECs showed fusiform shape or polygonal, regular cobblestone morphology after fusion to monolayer, and expressed factor VIII- related antigen (VIIIF - Ag ). Under fluorescence microscope, those cells demonstrated positive staining for uptaking acetylated low - density lipoprotein ( Dil - Ac - LDL ) and binding Bandeiraea simpli-cifolia lectin I ( FITC - BSI). The purity of PMVECs was above 90% . PMVECs formed capillary - like structures within 4 h to 6 h after plated on matrigel and fully developed a honeycomb appearance at 10 - 12 h, and then the capillary tubes began to break apart. CONCLUSION: The tissue block pasted method for primary cultivation of PMVECs is easy to handle, with high yield and purity, and will further facilitate related studies of well - established tube - formation assay in vitro.关键词
小鼠/肺微血管内皮细胞/原代培养/血管生成Key words
Mice/Pulmonary microvascular endothelial cells/Primary culture/Angiogenesis分类
医药卫生引用本文复制引用
高润娣,曹婕,卢珊,应琳,高璇,陆国华,李和权,周建英..小鼠肺微血管内皮细胞的培养鉴定及其血管形成功能的研究[J].中国病理生理杂志,2012,28(1):186-188,192,4.基金项目
浙江省自然科学基金资助项目(No.Y2100346) （No.Y2100346）
浙江省医药卫生科技计划项目(No.2008B080) （No.2008B080）
教育部博士点基金资助项目(No.20100101120119) （No.20100101120119）
