CAJ | 학술논문

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Ⅰ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.
목적 건립인자궁내막이위증원성성섬유세포(HEFC)적체외배양세포모형.방법 채용Ⅰ형、Ⅱ형화Ⅳ형효원매연합소화법종인란소자궁내막이위증낭종중분리성섬유세포,경밀도리심화차속첩벽법순화세포.광경화면역세포화학염색법대소획득적HEFC진행감정.결과 소획득적HEFC재면역세포화학염색분석중유성섬유세포적표지물파형단백적강렬표체,기섬유모세포적표지물α-SMA우이표체,상피세포적표지물각단백무표체;배양중적성섬유세포생장상태량호,정현중등장도적교관적사형、삼각형、성형화다각형;가량호적전대배양.결론 본연구건립료교이상적인HEFC체외배양체계,대료해인자궁내막이위증원성성섬유세포적이질성유중요개치,위종분자수평상연구인자궁내막이위증중섬유화급섬유점련적병리발생궤제제공료충족、가고적파세포.
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 specificities of HEFC.Sufficient and reliable target cells may be obtained for studying the mechanisms of fibrosis and adhesion in endometriosis at the molecular level.