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METHODS FOR ASSESSMENT OF PROLIFERATION IN AN IN VITRO TISSUE MODEL OF ORAL EPITHELIA.
Ackerman, G.L., Chlipata, E.
Premier Laboratory, LLC, University of Colorado, Boulder, CO.
Histologic, Vol. 38, No. 1, May 2005.
Keywords: EpiGingival, EpiOral, H&E, Histology (H&E staining), Immunohistochemical staining, Ki-67, Ki-67 nuclear protein, Ocular micrometer, Proliferation, Proliferative agents, Thickness
Summary:
This study demonstrated that a robust method for the assessment of the proliferative effects of agents applied to MatTek's EpiOral and EpiGingival in vitro tissue equivalents includes measurements of epithelial thickness, basal cell layer thickness, number of cells in the basal cell layer, and number of Ki-67 positive cells.
Introduction: EpiOral™ and EpiGingival™ cultures are organotypic in vitro models of oral and gingival epithelium developed by MatTek Corporation (Ashland, MA). Normal human oral keratinocytes are differentiated into tissues with a noncornified, buccal phenotype, or a cornified, gingival phenotype.
The same cells are used to propagate both tissues. The epithelium is cultured on a microporous membrane, Millipore Millicell® CM cell culture inserts (Millipore Corp., Billerica, MA) on a plastic disk utilizing serum-free media.
The model is used to assess the response of the epithelium to various stimuli including proliferative agents. Scientists at the University of Colorado and Premier Laboratory, LLC report here on methods used to assess proliferation in this model, including Ki-67 immunohistochemical staining, number of cells in the basal cell layer, thickness of the basal cell layer, and thickness of the epithelium.
Conclusion: Measurement of epithelial thickness, basal cell layer thickness, number of cells in the basal cell layer, and number of Ki-67 positive cells provides a robust method for assessment of proliferation in EpiOral and EpiGingival cultures.
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