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Development of a highly reproducible three-dimensional organotypic model of the oral mucosa

Nature Protocols volume 1, pages 2012–2018 (2006)Cite this article

Abstract

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.

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Figure 1
Figure 2: Histologic characterization of the 3D model of oral mucosa.
Figure 3: Application of the 3D model of the oral mucosa in the study of host–pathogen interactions.

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Acknowledgements

This study was supported by USPHS Research Grant RO1 DE13986 to ADB from the National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892.

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  1. Department of Oral Health and Diagnostic Sciences, School of Dental Medicine, University of Connecticut, 263 Farmington Ave, Farmington, 06030-1710, Connecticut, USA

    Anna Dongari-Bagtzoglou & Helena Kashleva

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  1. Anna Dongari-Bagtzoglou
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Correspondence to Anna Dongari-Bagtzoglou.

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Dongari-Bagtzoglou, A., Kashleva, H. Development of a highly reproducible three-dimensional organotypic model of the oral mucosa. Nat Protoc 1, 2012–2018 (2006). https://doi.org/10.1038/nprot.2006.323

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