中国组织工程研究2012，Vol.16Issue(15)：2691-2694,4.DOI:10.3969/j.issn.1673-8225.2012.15.007
改良贴壁法结合内皮细胞培养基培养小鼠肺微血管内皮细胞
Primary cultivation of mouse pulmonary microvascular endothelial cells by improved adherence method combined with endothelial cell culture medium
摘要
Abstract
BACKGROUND: Pulmonary microvascular endothelial cells (PMVECs) are one of the most important endothelial cell models for microcirculation. During many training methods, simple adherence method is relatively simple, but the time-consuming and more impurity cells are the biggest obstacle for cells culture. OBJECTIVE: To optimize the cultivation method for mouse PMVECs in vitro, and to observe the growth condition and to identify the property. METHODS: The pulmonary tissue particles were isolated from 5-day-old C57BL/6J mice pups by scissoring peripheral lung lobes accurately under sterile conditions, and cultured with endothelial cell medium. The morphology and growing characters of the cells were observed by inverted microscope. Cultured cells were identified by immunohistochemistry and immunofluorescence staining of factor related antigen. Ⅷ RESULTS AND CONCLUSION: A few spindle-shaped cells were found along the edge of tissue particles after cultured for 24 hours. After subculturing, cells spread even faster and took on a homogeneous cobblestone-like appearance, with the purity exceeding 98%. Morphological observations and detections of related antigen results conformed that these cultured cells were Ⅷendothelial cells. It is a convenient and effective approach to gain a large scale of PMVECs from neonatal mice by tissue-anchoring method and endothelial cell medium.关键词
肺微血管内皮细胞/细胞培养/优化/鉴定/小鼠/血管组织工程分类
医药卫生引用本文复制引用
黄偲璇,贺涛,邢怡桥..改良贴壁法结合内皮细胞培养基培养小鼠肺微血管内皮细胞[J].中国组织工程研究,2012,16(15):2691-2694,4.基金项目
国家自然科学基金项目青年科学基金(81000393),项目名称:15-LOX在缺氧导致的视网膜新生血管疾病中的作用及其机制. （81000393）
