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Three‐dimensional epithelial and mesenchymal cell co‐cultures form early tooth epithelium invagination‐like structures: Expression patterns of relevant molecules
Author(s) -
Xiao Li,
Tsutsui Takeki
Publication year - 2012
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.24056
Subject(s) - mesenchyme , matrigel , microbiology and biotechnology , mesenchymal stem cell , epithelium , cadherin , invagination , biology , epithelial–mesenchymal transition , pathology , anatomy , cell , downregulation and upregulation , medicine , biochemistry , genetics , gene
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, CD44 and E‐cadherin , and mesenchymal condensation involved molecules, N‐cadherin and Msx1 expressed 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 , and Pax9 are 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.