基础研究一种改良的肺微血管内皮细胞培养方法.pdfVIP

2012;32(8) 南方医科大学学报（J South Med Univ ） ·1151 · 基础研究 一种改良的肺微血管内皮细胞培养方法 1 2 1 1 1 1 胡国栋 ，陈英华 ，刘爱华 ，周国红 ，康 静 ，蔡绍曦 1 2 南方医科大学 南方医院呼吸科，基础医学院组织胚胎学教研室，广东 广州 510515 摘要：目的 对目前常用的小鼠肺微血管内皮细胞（PMVECs）原代培养方法进行改良，以期建立较为容易获取纯化的小鼠 PMVECs 的方法。方法 应用C57小鼠，采用改进的组织块贴壁法分离、培养PMVECs ，光镜观察细胞形态，Ⅷ因子相关抗原和 CD31 相关抗原免疫细胞化学法鉴定培养的PMVECs ，并比较改良培养方法与传统培养方法的优越性。结果体外培养的原代 PMVECs 在光镜下呈类圆形或短梭形，形成单层后呈铺路石样排列，Ⅷ因子相关抗原和CD31荧光染色阳性，利用改良的培养方 法培养的细胞生长较传统方法更加快速，分布更加均匀。结论改良的PMVECs 分离、培养方法获得的细胞生长状态良好，克服 细胞生长缓慢、杂细胞容易污染等难题，是一种较为理想的培养方法。 关键词：肺微血管内皮细胞；分离培养；鉴定 中图分类号：R-331 文献标志码：A 文章编号：1673-4254（2012）08-1151-03 doi: 10.3969/j.issn. 1673-4254.2012.08.17 /kcms/detail/44.1627.R1148.030.html An improved method for primary culture of mouse pulmonary microvascular endothelial cells 1 2 1 1 1 1 HU Guodong , CHEN Yinghua , LIU Aihua , ZHOU Guohong , KANG Jing , CAI Shaoxi 1Department of Respiratory Diseases, Nanfang Hospital, 2Department of Histology and Embryology, Southern Medical University, Guangzhou 510515, China Abstract: Objective To establish an improved method for culturing primary mouse pulmonary microvascular endothelial cells (PMVECs). Methods An improved tissue block adherent culture method was used to isolate and culture the PMVECs from C57 mice. The cultured cells were identified by factor VIII-related antigen and CD31 antigen, and the growth of cells cultured using the improved method and the conventional method was compared. Results The cultured primary pulmonary microvascular endothelial cells showed a short fusiform or round morphology, and the cell monolayer displayed a cobble stone-like appearance. The cultured cells were positive for VIII-related antigen