Lysis of whole blood in vitro causes DNA strand breaks in human lymphocytes

S Narayanan, M R O'Donovan, Susan Joyce Duthie

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

DNA damage in lymphocytes, as measured by alkaline single cell gel electrophoresis (pH 12.7), is greatly increased by the concurrent lysis of whole blood in both freshly isolated samples and in PHA-stimulated cultures over a period of 7 days. Further, there is a marked progressive increase in DNA damage with time in PHA-stimulated lymphocytes cultured in whole blood even when the lymphocytes are separated before analysis; no such increase is seen in lymphocytes cultured alone. This indicates that there are components in whole blood that can cause DNA damage in lymphocytes, with granulocytes and lysis of red blood cells likely candidates. The DNA damage is greatly reduced in granulocyte-depleted whole blood cultures, but even in these significant increases are seen at later sampling times. Consequently, careful sample preparation is of paramount importance if the Comet assay is to be successfully used to assess DNA damage in human peripheral blood lymphocytes. Further, the progressive increase in DNA damage in whole blood cultures may influence other methods using lymphocytes for population biomonitoring and may be significant for in vitro genotoxicity testing.

Original languageEnglish
Pages (from-to)455-459
Number of pages5
JournalMutagenesis
Volume16
Issue number6
Publication statusPublished - Nov 2001

Keywords

  • CELL GEL-ELECTROPHORESIS
  • COMET ASSAY
  • DAMAGE
  • CULTURES
  • STORAGE

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