Development of a highly reproducible three-dimensional organotypic model of the oral mucosa
Development of a highly reproducible three-dimensional organotypic model of the oral mucosa
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DOI：
10.1038/nprot.2006.323
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Kashleva, Helena
中科院分区：
文献类型：
--
作者：
Dongari-Bagtzoglou, Anna;Kashleva, Helena
在这份报告中，我们描述了一个标准化的三维（3D）系统的人口腔粘膜的基础上永生化的人口腔角质形成细胞系（OKF 6/TERT-2）的发展。该程序需要大约2-3周才能完成，包括三个主要阶段：胶原包埋的成纤维细胞的制备，粘膜成分的添加和培养物的空运，以确保充分的分化/分层。该过程导致多层上皮结构，其中层的组织类似于天然口腔粘膜中的细胞。具体地，该模型系统由具有一层柱状至圆形细胞的基底层、相对平坦的棘层和颗粒层以及非角质化角质层组成。该3D系统类似于基于细胞系TR 146（SkinEthic）的市售系统，不同之处在于我们的模型系统不包含角化不良变化并且具有粘膜下层组分，因此更好地代表正常人粘膜和粘膜下层。
In this report we describe the development of a standardized three-dimensional (3D) system of the human oral mucosa based on an immortalized human oral keratinocyte cell line (OKF6/TERT-2). The procedure takes approximately 2-3 weeks to complete and includes three main stages: preparation of collagen-embedded fibroblasts, addition of the mucosal component and airlifting of cultures to ensure adequate differentiation/stratification. This procedure results in a multilayer epithelial structure in which layers are organized similarly to the cells in native oral mucosa. Specifically, this model system consists of a stratum basale, having one layer of columnar to round cells, a relatively flattened stratum spinosum and stratum granulosum, and a non-keratinizing stratum corneum. This 3D system resembles the commercially available system based on the cell line TR146 (SkinEthic), with the exception that our model system does not contain dyskeratotic changes and has a submucosal component, and thus better represents the normal human mucosa and submucosa.