人子宫旁韧带成纤维细胞原代培养的初步研究  

Preliminary Study on Primary Culture of Human Parametrial Ligament Fibroblasts

作　　者：丁文娟[1] 洪莎莎[1] 闵洁[1] 方桂[1] 张小红[1] 胡鸣[1] 杨青[1] 赵杨[1] 洪莉[1] 

机构地区：[1]武汉大学人民医院,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2014年第1期16-19,共4页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81270684);湖北省卫生厅资助项目(编号:NX2011-5);武汉市科技局资助项目(编号:201260523178-1)

摘　　要：目的:初步探索和建立人子宫旁韧带成纤维细胞原代培养的方法,为盆腔器官脱垂的基础研究提供成熟的体外人盆底成纤维细胞系。方法:临床搜集因盆腔器官脱垂(POP)或非POP的良性妇科疾病而行子宫切除患者的子宫旁韧带(包括主韧带和骶韧带),组织剪碎后采用改良的Ⅰ型胶原酶和胰蛋白酶消化法进行原代培养,待细胞长至贴满培养瓶底面80%以上时,消化传代,对所培养的成纤维细胞进行形态学观察及免疫荧光鉴定其来源。结果:原代培养第3-4天开始,可见有单个成纤维细胞迁出、生长,7-10d时可见少部分细胞逐渐贴壁形成较小的成纤维细胞团,第25天左右多个细胞团生长中心爬出的成纤维细胞互相连接聚集成片,铺满培养瓶底;采用免疫组化染色进行细胞鉴定,细胞波形蛋白表达阳性,角蛋白表达阴性,证实其为成纤维细胞。结论:本研究确立了体外人子宫旁韧带成纤维细胞原代培养,为后续研究POP发病机制提供成熟的体外宫旁韧带成纤维细胞。Objective： To preliminarily establish primary culture methods and provide mature cell lines on human parametrial ligament fibroblasts. Methods. Specimen were taken from part of the para- metrial ligaments （including sacral and cardinal ligaments） by surgery in patients with or without pelvic organ prolapse （POP） who underwent hysterectomy surgery. The morphological changes were observed under microscope and identified by immunohistochemistry. Results： Single fibro- blast began to grow and move out in 3 to 4 days of primary culture, small groups of fibroblasts gradually were formed in 7 to 10 days, and on the 25th day the cells were connected to each other and formed a network structure. The vimentin was positively expressed by immunohistochemical staining, while keratin （cytokeratin） was negatively detected. Conclusion： The study cultured mature human parametrial ligament fibroblasts for the next study.

关 键 词：人子宫旁韧带 成纤维细胞 原代培养 

分 类 号：R711.23[医药卫生—妇产科学]