DNA damage in oral mucosal epithelial cells cultured in complex and xenobiotic-free media: a comparison study

In this study, we evaluated the genomic stability of oral mucosal epithelial cells (OMECs) cultured in complex media (COM) and xenobiotic-free media (XF) to assess their potential clinical application for limbal stem cell deficiency (LSCD) treatments. OMECs serve as a promising autologous cell source for bilateral LSCD treatment, offering an alternative to limbal epithelial cells (LECs). However, genomic integrity is crucial to ensure the long-term success of transplanted cells. We performed micronucleus (MNi) tests and comet assays to compare DNA damage in OMECs cultured in both media types. The results indicated no significant differences in cell morphology, viability, or size between the two conditions. The MNi frequency was similar, with 5.67 and 6.17 MNi per 1,000 cells in COM and XF conditions, respectively. Comet assay results showed low levels of strand breaks (SBs) and oxidized DNA lesions in both media, with XF showing a slightly lower, albeit statistically insignificant, percentage of tail DNA for net Fpg-sensitive sites. Our findings suggest that OMECs can be effectively cultivated in either COM or XF media without inducing significant DNA damage, supporting the potential use of XF media in clinical settings to reduce contamination risks. This study underscores the importance of genomic stability in cultured cells for ocular surface transplantation, contributing valuable insights into optimizing culture conditions for safer and more effective clinical applications.