| Autor: |
Nora, Markanović, Darija, Ibrahimagić, Lejla, Čaluk Klačar, Irma, Durmišević, Tamara, Ćetković Pećar, Anja, Haverić |
| Předmět: |
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| Zdroj: |
Genetics & Applications; 2023 Special Issue, p39-39, 1p |
| Abstrakt: |
The comet assay is a simple and versatile method widely used in the assessment of DNA damage, such as DNA single-strand breaks and alkali-labile sites in DNA nucleoids. Adherent cell lines are often used in assessment of cytotoxicity and genotoxicity, but detachment procedure may be confounder. Trypsin is the most common agent used for detachment; however it can damage cells and contribute to the elevated levels of DNA damage. In our studies, we assessed trypsin-induced DNA damage over different time periods (1, 2, 4 and 6 minutes) in two cell lines: MDBK (Madin-Darby bovine kidney) and human bladder carcinoma 5637. The aim was to determine the optimal period of trypsin treatment that would result in minimal DNA damage, and efficient detachment yield. Results revealed the highest trypsininduced DNA damage in 1-minute treatment of MDBK cells; significantly increased compared to 2 and 4-minutes treatments. This suggests resistance of MDBK cells to trypsin-induced damage, and their efficient repair mechanisms. In 5637 cell line significant and positive association was found between the length of treatment period and the level of DNA damage, showing that this cell line is sensitive to trypsin treatment. Trypsin treatment optimization for each used cell line is highly needed for DNA damage analysis. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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