Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Comparison of Culture Media for in-vitro Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Version 1 : Received: 4 January 2023 / Approved: 9 January 2023 / Online: 9 January 2023 (07:35:28 CET)
Version 2 : Received: 30 March 2023 / Approved: 3 April 2023 / Online: 3 April 2023 (03:33:37 CEST)

A peer-reviewed article of this Preprint also exists.

Cuadra, G.A.; Shamim, A.; Shah, R.; Morgan, J.; Palazzolo, D.L. Comparison of Culture Media for In Vitro Expansion of Oral Epithelial Keratinocytes. Appl. Biosci. 2023, 2, 308-327. Cuadra, G.A.; Shamim, A.; Shah, R.; Morgan, J.; Palazzolo, D.L. Comparison of Culture Media for In Vitro Expansion of Oral Epithelial Keratinocytes. Appl. Biosci. 2023, 2, 308-327.

Abstract

Background: Understanding in vitro expansion of OKF6/TERT-2 oral epithelial cells is important for studying molecular biology of disease and pathology affecting the oral cavity. The media used for any cell culture is paramount in terms of efficient output. Therefore, this study aims to compare two different media for OKF6/TERT-2 cultures: Keratinocyte Serum-Free Medium (KSFM) and a composite medium comprised of DMEM/F-12 mixed with KSFM (referred to as DFK). For application purposes, this investigation also compares the toxicological effects of flavored electronic cigarette liquids (E-liquids) on OKF6/TERT-2 cultures grown in both media. Methods: Cells were grown in KSFM and DFK media and cellular growth, morphology, gene expression of mucins and tight junctions, as well as wound-healing were determined. Additionally, cellular viability and cytotoxicity were indexed after E-liquid exposures. Results: While overall cellular morphologies remained unaltered, cells grown in DFK reached confluency faster. Except for claudin-1, there is no appreciable difference in expression of the other genes tested. Additionally, cultures in DFK appear more sensitive to E-liquids ± flavors. Conclusions: DFK is an alternative medium for cultivation of OKF6/TERT-2 cells to study molecular biology of disease and pathology, such as their responses to E-liquids ± flavors.

Keywords

oral; mucosa; mucins; tight junctions; wound-healing; E-liquids; cytotoxicity; viability; confluency

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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