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DNA damage determined by Single cell gel electrophoresis using cryopreserved lymphocyte, frozen whole blood in EDTA tube or DNA stabilized tube
Author(s) -
Ronchi Carlos Fernando,
BerchieriRonchi Carolina Bragiola,
Correa Camila Renata,
Ferreira Ana Lucia dos Anjos,
Zhao Yan,
Kim Sung Woo,
Yeum KyungJin
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.535.2
Subject(s) - comet assay , cryopreservation , dna damage , whole blood , dna , microbiology and biotechnology , gel electrophoresis , chemistry , lymphocyte , andrology , biology , biochemistry , immunology , medicine , genetics , embryo
Oxidative DNA damage is of great importance due to the growing recognition that such damage can initiate and promote chronic diseases. To determine the usefulness of frozen whole blood on the analysis of DNA damage, lymphocytes isolated from the fresh blood followed by cryopreservation as well as collected from the frozen whole blood were analyzed in parallel for DNA damage. Lymphocytes were isolated from fresh blood from sows (n=10) and cryopreserved (Cryo). Whole blood samples were also collected in EDTA‐tubes (EDTA) as well as DNA‐stabilized tubes (DNA). Cryopreserved lymphocytes as well as lymphocytes recovered from the frozen whole blood were analyzed by alkaline single cell gel electrophoresis (comet assay) to determine the single and double strand brakes. The comet assay indicated less DNA damage (% damage) in the lymphocytes obtained from the frozen whole blood in DNA‐tube as compare to those of cryopreserved lymphocyte (Cryo, 45.8 ± 5.88 > DNA, 37.52 ± 6.72 = EDTA, 40.1 ± 3.65, p<0.05). Similar results obtained when the DNA was challenged with H 2 O 2 (Cyro, 51.9 ± 2.04 > DNA, 43.3 ± 5.43 = EDTA, 40.7 ± 6.42, p<0.05). The current study indicates that frozen whole blood can be used for DNA damage analysis and DNA‐preservation tube shows better response to the oxidative stress challenge. The presence of antioxidants in the whole blood may have a protective effect against oxidative DNA damage that might occur during the storage, which warrants further study. [Supported by CAPES0635‐09, FAPESP2008/08199‐2, USDA 1950‐51000‐065‐08S]