Cabral Joao Victor, Vodenkova Sona, Tomasova Kristyna, Vodickova Ludmila, El Yamani Naouale, Rundén Pran Elise, Dusinska Maria, Safanda Adam, Jirsova Katerina
Laboratory of Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, 1st Faculty of Medicine and General Teaching Hospital, Charles University, Czech Republic.
Department of Molecular Biology of Cancer, Institute of Experimental Medicine of the Czech Academy of Sciences, Prague, Czech Republic.
Mutagenesis. 2025 Aug 29;40(3):526-532. doi: 10.1093/mutage/geaf008.
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.
在本研究中，我们评估了在复合培养基（COM）和无外源化合物培养基（XF）中培养的口腔黏膜上皮细胞（OMECs）的基因组稳定性，以评估其在角膜缘干细胞缺乏症（LSCD）治疗中的潜在临床应用价值。OMECs是双侧LSCD治疗中一种有前景的自体细胞来源，可作为角膜缘上皮细胞（LECs）的替代物。然而，基因组完整性对于确保移植细胞的长期成功至关重要。我们进行了微核（MNi）试验和彗星试验，以比较在两种培养基中培养的OMECs的DNA损伤情况。结果表明，两种培养条件下细胞的形态、活力或大小均无显著差异。微核频率相似，在COM和XF条件下，每1000个细胞中的微核数分别为5.67和6.17个。彗星试验结果显示，两种培养基中的链断裂（SBs）和氧化DNA损伤水平均较低，XF中对Fpg敏感位点的净尾DNA百分比略低，尽管在统计学上无显著差异。我们的研究结果表明，OMECs可以在COM或XF培养基中有效培养，而不会诱导显著的DNA损伤，这支持了在临床环境中使用XF培养基以降低污染风险的可能性。本研究强调了培养细胞的基因组稳定性在眼表移植中的重要性，为优化培养条件以实现更安全、更有效的临床应用提供了有价值的见解。
