Isolamento e análise de indicadores genotóxicos em célulastronco de polpa dentária de cães

Abstract

The double-strand DNA breaks are considered one of the most lethal forms of damage that can compromise the ability of stem cells self-renew and differentiate. Therefore, additional studies are necessary to identify genome damage biomarkers that help the researchers to choice the better cell line to apply in clinical use. Here, we evaluated the potential use of DNA Comet Assay, DNA fragmentation fluorimetric assay using Picogreen dye and reactive oxygen species (ROS) level by DCFH-DA fluorimetric assay as potential biomarkers to identify the genome conditions of dental stem cells (DSCs) isolated from dog´s canine teeth. The results showed important differences among the three initial DSCs passage when compared to > 4th passages. In general, the initial DSCs passages presented higher cell proliferation, lower DNA damage and ROS. However, we noticed a large number of nucleus with some level of DNA damage (30 to 40% in the initial DSCs passage and > 50% in the late passage) that indicates an in vitro DSCs genomic fragility. The results suggest that these relatively simple and inexpensive approaches as Comet, DNA fragmentation could to help for sorting stem cell with less DNA damage used to research or therapeutic use.