Ouyang Zhuo, Liang Yuan-jing, Zhang Wei-yuan, Lu Dan
Department of Gynecology, Beijing Obstetrics & Gynecology Hospital, Capital Medical University, Beijing 100026, China.
Zhonghua Yi Xue Za Zhi. 2011 May 10;91(17):1207-10.
To establish the model of cultivating and identifying fibroblast from human endometriosis (HEFC) in vitro.
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.
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, star-shaped and polygonal fiber-like cells. Furthermore HEFC could be well sub-cultured.
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.
建立体外培养和鉴定人子宫内膜异位症成纤维细胞(HEFC)的模型。
采用Ⅰ型、Ⅱ型和Ⅳ型胶原酶消化人卵巢子宫内膜异位囊肿组织,通过离心和差速贴壁法纯化所得细胞。在倒置显微镜下观察形态变化鉴定HEFC,并采用免疫细胞化学法检测波形蛋白、α -平滑肌肌动蛋白(α-SMA)和角蛋白的表达。
培养的成纤维细胞波形蛋白免疫组化染色呈阳性,α-SMA极少呈阳性,角蛋白完全呈阴性。HEFC生长为短胖梭形、三角形、星形和多边形纤维样细胞的汇合单层。此外,HEFC可顺利传代培养。
获得的成纤维细胞可在体外稳定培养。研究HEFC的特性非常重要。可为在分子水平研究子宫内膜异位症的纤维化和粘连机制获得充足且可靠的靶细胞。