兔声带成纤维细胞的体外分离培养、鉴定与标记  

In vitro culture,identification and label of rabbit vocal cord fibroblasts

作　　者：牛玉光[1] 黄冬雁[1] 王荣光[1] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科,北京100853

出　　处：《军医进修学院学报》2010年第6期589-590,F0003,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：国家自然科学基金项目(30700937)~~

摘　　要：目的探讨体外培养兔声带成纤维细胞(Vocal Fold Fibroblasts,VFFs)的培养及标记方法。方法消化培养法分离、培养兔声带成纤维细胞,倒置显微镜观察其生长和形态特征,并传代、鉴定。增强型绿色荧光蛋白腺病毒载体(Ad-EGFP)标记体外培养的VFFs,取12只新西兰兔,以1×100万/0.1ml分别注入损伤模型左侧声带,观察注入VFFs的生长情况。结果兔声带成纤维细胞可以在体外原代和传代培养,具有成纤维细胞的典型形态。波形蛋白免疫荧光染色阳性,Ad-EFGP可以成功转染VFFs。将VFFs诸如声带后15d,组织学切片未发现成活细胞。结论用消化培养法可成功获得VFFs,Ad-EGFP是一种理想的示踪标记物。Objective To investigate the best method to culture and label vocalfold fibroblasts（VFFs）in vitro.Methods VFFs were cultured by enzyme digestion to observe their growth and morphology under an inverted microscope and labeled with Ad-EGFP.The cells（100 million/0.1ml）were injected into the vocal cords of 12 New Zealand rabbits to observe the growth of VFFs.Results VFFs could be cultured in vitro and showed characteristic morphologyof fibroblasts.Immunochemistry showed positive vimentin expression Ad-EGFP could transfect VFFs.No survived cells were observed on the tissue sections 15 days after injection of VFFs.Conclusion VFFs can be cultured by enzyme digestion.Ad-EGFP is an ideal tracing marker.

关 键 词：声带 成纤维细胞 兔 

分 类 号：R767.4[医药卫生—耳鼻咽喉科]