摘要
目的建立人子宫内膜异位症源性成纤维细胞（HEFC）的体外培养细胞模型。方法采用l型、Ⅱ型和Ⅳ型胶原酶联合消化法从人卵巢子宫内膜异位症囊肿中分离成纤维细胞，经密度离心和差速贴壁法纯化细胞。光镜和免疫细胞化学染色法对所获得的HEFC进行鉴定。结果所获得的HEFC在免疫细胞化学染色分析中有成纤维细胞的标志物波形蛋白的强烈表达，肌纤维母细胞的标志物α-SMA偶尔表达，上皮细胞的标志物角蛋白无表达；培养中的成纤维细胞生长状态良好，呈现中等长度的较宽的梭形、三角形、星形和多角形；可良好的传代培养。结论本研究建立了较理想的人HEFC体外培养体系，对了解人子宫内膜异位症源性成纤维细胞的异质性有重要价值，为从分子水平上研究人子宫内膜异位症中纤维化及纤维粘连的病理发生机制提供了充足、可靠的靶细胞。
Objective To establish the model of cultivating and identifying fibroblast from human endometriosis （HEFC） in vitro. Methods The tissues of human endometriotic cysts of ovary were digested by collagenases I , II and Ⅳ The resulting cells were purified by centrifugation and differential adhesion. HEFC was identified by observing the morphologic changes under an inverted microscope and the expressions of vimentin,α-SMA （α-smooth muscle actin）and keratin were detected by immunocytochemistry. Results Immunohistochemical staining of vimentin was positive, α-SMA rarely positive and keratin completely negative in cultured fibroblasts. HEFC grew as a confluent monolayer of short fat fusiform, triangular, starshaped and polygonal fiber-like cells. Furthermore HEFC could be well sub-cultured. Conclusion Acquired fibroblast can be cultured in vitro stably. It is quite important to study the speeificities of HEFC. Sufficient and reliable target cells may be obtained for studying the mechanisms of fibrosis and adhesion in endometriosis at the molecular level.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2011年第17期1207-1210,共4页
National Medical Journal of China
基金
2010年首都医科大学基础临床合作基金（10JL59）
