Three-dimensional epithelial and mesenchymal cell co-cultures form early tooth epithelium invagination-like structures : expression patterns of relevant molecules
Three-dimensional epithelial and mesenchymal cell co-cultures form early tooth epithelium invagination-like structures : expression patterns of relevant molecules
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三维上皮和间充质细胞共培养形成早期牙上皮内陷样结构：相关分子的表达模式
DOI：
10.1002/jcb.24056
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发表时间：
2012
期刊：
影响因子：
4
通讯作者：
Takeki Tsutsui
中科院分区：
文献类型：
--
作者：
Li Xiao;Takeki Tsutsui
上皮内陷是早期牙齿发育的关键特征。在这项研究中，我们建立了一个三维（3D）模型来通过组织工程来表示上皮内陷样结构。人正常口腔上皮细胞（OEC）和牙髓干细胞（DPSC）在基质胶或胶原凝胶上共培养2-7周，形成上皮和间质组织。通过HE染色、免疫染色和定量实时RT-PCR（qRT-PCR）分析组织学变化和基因表达。培养 4 周后，OEC 形成的上皮内陷到基质胶和胶原凝胶上的 DPSC 衍生间充质中。 OEC-DPSC 在基质胶上的共培养显示出典型的上皮细胞内陷和下面的间充质细胞的浓缩。上皮内陷相关分子CD44和E-cadherin以及间质凝结相关分子N-cadherin和Msx1在组织模型中高水平表达，表明上皮内陷是功能性的。然而，当OEC和DPSC在胶原凝胶上共培养时；经过长时间的培养，内陷的上皮转化为间充质内的多个上皮集落。当 DPSC 与永生化人类 OEC NDUSD-1 共培养时，所有上述特征均未呈现。免疫组织学染色和 qRT-PCR 分析表明，p75、BMP2、Shh、Wnt10b、E-钙粘蛋白、N-钙粘蛋白、Msx1 和 Pax9 参与 3D OEC-DPSC 共培养物中上皮内陷和上皮间质相互作用的启动。我们的结果表明，在一定条件下在基质胶上共培养 OEC 和 DPSC 可以建立上皮内陷样模型。该模型可能被探索作为上皮间质相互作用和牙齿再生的潜在研究工具。 J.细胞。生物化学。 113：1875–1885，2012。© 2012 Wiley periodicals, Inc.
Epithelium invagination is the key feature of early tooth development. In this study, we built a three‐dimensional (3D) model to represent epithelium invagination‐like structure by tissue engineering. Human normal oral epithelial cells (OECs) and dental pulp stem cells (DPSCs) were co‐cultivated for 2–7 weeks on matrigel or collagen gel to form epithelial and mesenchymal tissues. The histological change and gene expression were analyzed by HE staining, immunostaining, and quantitative real‐time RT‐PCR (qRT‐PCR). After 4 weeks of cultivation, OECs‐formed epithelium invaginated into DPSCs‐derived mesenchyme on both matrigel and collagen gel. OEC–DPSC co‐cultures on matrigel showed typical invagination of epithelial cells and condensation of the underlying mesenchymal cells. Epithelial invagination‐related molecules,CD44andE‐cadherin, and mesenchymal condensation involved molecules,N‐cadherinandMsx1expressed at a high level in the tissue model, suggesting the epithelial invagination is functional. However, when OECs and DPSCs were co‐cultivated on collagen gel; the invaginated epithelium was transformed to several epithelial colonies inside the mesenchyme after long culture period. When DPSCs were co‐cultivated with immortalized human OECs NDUSD‐1, all of the above‐mentioned features were not presented. Immunohistological staining and qRT‐PCR analysis showed that p75,BMP2,Shh,Wnt10b,E‐cadherin,N‐cadherin,Msx1, andPax9are involved in initiating epithelium invagination and epithelial–mesenchymal interaction in the 3D OEC–DPSC co‐cultures. Our results suggest that co‐cultivated OECs and DPSCs on matrigel under certain conditions can build an epithelium invagination‐like model. This model might be explored as a potential research tool for epithelial–mesenchymal interaction and tooth regeneration. J. Cell. Biochem. 113: 1875–1885, 2012. © 2012 Wiley Periodicals, Inc.