Culturing Human Oral Keratinocytes and Gingival Epithelium

Cell cultures: The human oral keratinocyte cell line OKF6/TERT-1 was grown in keratinocyte serum free medium (KSFM) supplemented with L-glutamine and penicillin-streptomycin-fungizone (Sigma-Aldrich) in the presence of 0.03 M calcium chloride and bovine pituitary extract as described previously (19 (link)). Three-dimensional cultures of normal human gingival epithelial cells were obtained from MatTek. These represent confluent, terminally differentiated, multi-layer primary cultures of ginigival epithelium (EpiGingiva), obtained in 24-well transwell plates obtained from the producer (Supplementary Figure S1). After delivery, the transwells are transferred to 6-well plates into the medium provided, and the cultures are maintained in an air-liquid interface for 1–2 days prior to their use in the experiment.

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Figgins E.L., Arora P., Gao D., Porcelli E., Ahmed R., Daep C.A., Keele G., Ryan L.K, & Diamond G. (2024). Enhancement of innate immunity in gingival epithelial cells by vitamin D and HDAC inhibitors. Frontiers in Oral Health, 5, 1378566.

Publication 2024

penicillin-streptomycin-fungizone

Corresponding Organization :

Other organizations : Florida College, University of Florida, University of Louisville, Colgate-Palmolive (United States)

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Variable analysis

independent variables

Cell lines used: OKF6/TERT-1 (human oral keratinocyte cell line) and normal human gingival epithelial cells (three-dimensional cultures)

dependent variables

Growth and differentiation of the cell cultures

control variables

Culture medium: keratinocyte serum free medium (KSFM) supplemented with L-glutamine and penicillin-streptomycin-fungizone
Presence of 0.03 M calcium chloride and bovine pituitary extract in the medium
Maintenance of the gingival epithelial cell cultures in an air-liquid interface for 1-2 days prior to the experiment

controls

No positive or negative controls were explicitly mentioned in the provided information.

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