Isolation of leukocytes from frozen buffy coat for comet assay analysis of DNA damage
| Autor: | Sergey Shaposhnikov, Siv Kjølsrud Bøhn, Andrew Collins, Vaineta Vebraite |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Adult Guanine DNA damage Health Toxicology and Mutagenesis Buffy coat Cell Separation 010501 environmental sciences 01 natural sciences Peripheral blood mononuclear cell 03 medical and health sciences Genetics Centrifugation Density Gradient Leukocytes Humans 0105 earth and related environmental sciences Cryopreservation Chemistry DNA Breaks DNA Hydrogen Peroxide Middle Aged Isolation (microbiology) Molecular biology Comet assay 030104 developmental biology DNA-Formamidopyrimidine Glycosylase Blood Preservation Blood Buffy Coat Leukocytes Mononuclear Percoll gradient centrifugation Female Comet Assay DNA Damage |
| Zdroj: | Mutation research. Genetic toxicology and environmental mutagenesis. 843 |
| ISSN: | 1879-3592 |
| Popis: | Frozen buffy coat fractions are often stored in human biomonitoring trials but their use for biomarker purposes has been limited. The purpose of the current study was to study whether frozen buffy coats can be used to monitor DNA damage levels. EDTA blood samples were provided from 9 healthy, non-smoking female volunteers, aged 26–48. Pre-existing DNA damage (strand breaks and oxidised purines) was measured with the comet assay in thawed resuspended buffy coat samples and washed leukocytes from these buffy coats, as well as resistance to DNA damage induced exogenously by H2O2 in the latter, and compared with damage measured in peripheral blood mononuclear cells isolated from fresh blood using percoll gradient centrifugation. Basal DNA damage levels (strand breaks) were significantly higher in the leukocytes isolated from frozen buffy coats in the untreated samples compared with peripheral blood mononuclear cells. However, the levels of strand breaks were still low ( The results indicate that buffy coat samples stored from cohort studies might be usefully analysed for DNA damage in retrospective epidemiological investigations. However, caution should be exercised when comparing the absolute levels of DNA damage in buffy coat leukocytes and peripheral blood mononuclear cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect