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作 者:胡国栋[1] 陈英华[2] 刘爱华[1] 周国红[1] 康静[1] 蔡绍曦[1]
机构地区:[1]南方医科大学南方医院呼吸科 [2]南方医科大学基础医学院组织胚胎学教研室,广东广州510515
出 处:《南方医科大学学报》2012年第8期1151-1153,共3页Journal of Southern Medical University
基 金:国家973计划项目(2012CB518203);广东省科技计划项目(2011B031800245);广东省医学科研基金(B2012218);南方医院院长基金(2010c002;2011z005)~~
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。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 and CD31 antigen. The cell growth was accelerated in the cell cultures with the improved method compared with that in conventional cell cultures. Conclusion The improved culture method allows more efficient acquisition of primary mouse PMVECs of a greater purity.
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