In vitro three-dimensional modelling of human ovarian surface epithelial cells
In vitro three-dimensional modelling of human ovarian surface epithelial cells
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DOI：
10.1111/j.1365-2184.2009.00604.x
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发表时间：
2009-06-01
影响因子：
8.5
通讯作者：
Dafou, D.
中科院分区：
文献类型：
--
作者：
Lawrenson, K.;Benjamin, E.;Dafou, D.
90%的恶性卵巢癌是上皮性的，被认为是由卵巢表面上皮（OSE）引起的。我们假设，原代OSE细胞的生物学特性将更接近OSE在体内，如果建立为三维（3D）culture.OSE细胞培养的多细胞球体（MCS）（i）在旋转细胞培养系统（RCCS）和（ii）polyHEMA涂层塑料。通过电子显微镜检查MCSs，并将其与2D中生长的原代组织和细胞的OSE进行比较。Annexin V FACS分析用于评估细胞凋亡，细胞外基质（ECM）蛋白的表达通过免疫组织化学染色进行分析。与polyHEMA MCSs相比，RCCS MCSs结构紊乱，凋亡细胞比例更高，并且相同的细胞在2D培养中生长。在2D中，AE 1/AE 3，层粘连蛋白和波形蛋白的广泛表达是不可检测的免疫组化，而这些蛋白质的强表达被观察到在相同的细胞生长在3D文化和OSE的原代tissues.Physiological和生物学特性的OSE细胞生长在3D文化更接近的特点OSE细胞在体内比经典的2D方法生长。建立体外3D OSE模型将有助于更好地了解上皮性卵巢癌的肿瘤转化机制。
Ninety percent of malignant ovarian cancers are epithelial and thought to arise from the ovarian surface epithelium (OSE). We hypothesized that biological characteristics of primary OSE cells would more closely resemble OSE in vivo if established as three-dimensional (3D) cultures.OSE cells were cultured as multicellular spheroids (MCS) (i) in a rotary cell culture system (RCCS) and (ii) on polyHEMA-coated plastics. The MCSs were examined by electron microscopy and compared to OSE from primary tissues and cells grown in 2D. Annexin V FACS analysis was used to evaluate apoptosis and expression of extracellular matrix (ECM) proteins was analysed by immunohistochemical staining.On polyHEMA-coated plates, OSE spheroids had defined internal architecture. RCCS MCSs had disorganized structure and higher proportion of apoptotic cells than polyHEMA MCSs and the same cells grown in 2D culture. In 2D, widespread expression of AE1/AE3, laminin and vimentin were undetectable by immunohistochemistry, whereas strong expression of these proteins was observed in the same cells grown in 3D culture and in OSE on primary tissues.Physiological and biological features of OSE cells grown in 3D culture more closely resemble characteristics of OSE cells in vivo than when grown by classical 2D approaches. It is likely that establishing in vitro 3D OSE models will lead to greater understanding of the mechanisms of neoplastic transformation in epithelial ovarian cancers.