人主动脉瓣间质细胞的分离、培养、鉴定及表型分析  

Isolation,culture,identification and phenotypic analysis of human aortic valve interstitial cells

作　　者：江磊[1] 徐平[1] 高洪波[1] 郝传吉[1] 黄强[1] 王伟[1] 于仁斌[1] 

机构地区：[1]青岛大学医学院附属医院心血管外科,山东青岛266003

出　　处：《心脏杂志》2009年第2期206-208,共3页Chinese Heart Journal

摘　　要：目的建立人主动脉瓣间质细胞体外培养方法并鉴定其形态、类型及膜表面分子分布。方法将瓣膜置换术中获取的畸形瓣膜及风心病瓣膜内皮层剥除,用I型胶原酶消化后铺养。光镜下观察细胞的形态,将其以抗人α-平滑肌肌动蛋白抗体及核染色双标记后,以激光共聚焦显微镜观察细胞的类型。以抗干扰素(IFN)-γ受体单抗及抗肿瘤坏死因子(TNF)-α受体单抗标记细胞后,用流式细胞术分析细胞膜分子的分布。结果分离培养的主动脉间质细胞主要为肌成纤维细胞。畸形瓣膜和风心病瓣膜表面IFN-γ受体及TNF受体的分布状态不同。结论成功地建立了体外培养瓣膜间质细胞的方法,并发现来源不同的间质细胞膜表面分子的分布迥异。AIM To isolate and culture human aortic valve interstitial cells and to identify their morphology, subtype and membrane molecules distribution. METHODS The endothelial cell layers on the dissected valves were removed and the remaining interstitial layers were minced and digested with collagenase I. Cells were then seeded and maintained on culture dishes. Cells were observed under light microscopy and cellular α-smooth muscle actin was detected with confocal staining microscopy. The expressed interferon （IFN）-γ and tumor necrosis factior（TNF）-α receptors were analyzed by flow cytometry with their corresponding monoclonal antibodies. RESULTS The isolated valves interstitial cells were proved to be myo- fibroblast. More IFN-γ, receptors and TNF-α receptors were expressed on rheumatic aortic valves interstitial cells than on non-rheumatic cells and their expression was patients＇ sera dependent. CONCLUSION The valves interstitial cells culture system has been established successfully and the membrane molecules differ according to their origins.

关 键 词：主动脉瓣 问质细胞 肌成纤维细胞 先天畸形 风心病 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学]