摘要
目的 探讨一种体外分离、扩增人主动脉瓣膜间质细胞的方法并进行表型鉴定,建立研究主动脉瓣疾病的体外实验细胞模型.方法 选取在北京协和医院接受手术治疗的急性Stanford A型主动脉夹层患者5例,其中男性3例,女性2例,年龄43～ 55岁,平均（49.4±4.5）岁,术中取其正常主动脉瓣膜组织,采用改进的胶原酶消化法分离并体外扩增人主动脉瓣膜间质细胞,对原代培养细胞采用波形蛋白（Vimentin）染色及α-平滑肌肌动蛋白（α-SMA）染色进行表型鉴定.结果 改进的胶原酶消化法可成功分离并体外扩增瓣膜间质细胞,体外原代培养的人主动脉瓣膜间质细胞,波形蛋白（Vimentin）和α-平滑肌肌动蛋白（α-SMA）免疫荧光染色阳性.结论 改进的胶原酶消化法可成功建立人主动脉瓣膜间质细胞体外实验细胞模型,细胞表型鉴定证实其达到实验要求,从而为从细胞水平研究退行性主动脉瓣疾病的发病机制奠定了基础.
Objective To explore a method to culture human aortic valvular interstitial cells and identify the phenotypes,to establish the cell model which would be used to study aortic valve diseases in vitro.Methods Normal aortic valves of the patient with acute Stanford A aortic dissection in Peking Union Medical College Hospital were preserved during the surgical operation.Human aortic valvular interstitial cells were isolated and amplified in vitro by modified collagenase digestion method.The cell phenotype was identified by the immunofluorescent staining.Results Human aortic valvular interstitial cells could be successfully isolated and amplified in vitro by modified collagenase digestion method,identified by positive staining of Vimention and α-SMA.Conclusions The cell model of human aortic valvular interstitial ceils could be successfully established in vitro by modified collagenase digestion method.The cell phenotype identification proved to meet the experimental requirements.So it could provide cellular foundations for the study of pathogenesis of degenerative aortic valve disease.
出处
《国际外科学杂志》
2015年第6期393-395,F0003,共4页
International Journal of Surgery
关键词
人类
主动脉瓣
间质细胞
细胞培养
表型
Humans
Aortic valve
Interstitial cell
Cell culture techniques
Phenotype
作者简介
通信作者：苗齐，Email：miaoqipumc@hotmail．com
