Effects in the Comet assay of storage conditions on human blood.

Author(s): Anderson D, Yu TW, Dobrzynska MM, Ribas G, Marcos R

Publication: Teratog Carcinog Mutagen, 1997, Vol. 17, Page 115-25

PubMed ID: 9436261 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage temperature and duration on human lymphocyte cell viability and DNA damage.

   Summary of Findings:

   Storage of lymphocytes at 4 degrees C or room temperature did not affect cell viability. For the most part, storage for up to 4 days at room temperature or 4 degrees C did not significantly alter comet assay outcomes in untreated cells, however after 7 days of storage at either temperature, outcomes were significantly different. Some statistically significant changes in comet assay outcomes were observed after treatment with BLM or ENU when compared to no treatment, however these changes were variable when storage duration was taken into account, and no trends were observed. The authors state that the changes were not biologically significant. Storage at -20 degrees C without cryopreservative caused significant increases in DNA damage, with or without BLM or ENU treatment.

   Biospecimens

   • Cell - White Cells

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Light microscopy
   DNA	Comet assay

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	Room temperature (22-29 degrees C) 4 degrees C -20 degrees C
   Storage	Storage duration	0 d 1 d 2 d 3 d 4 d 7 d
   Biospecimen Aliquots and Components	Blood and blood products	Lymphocyte
   Comet assay Specific	Reaction solution	PBS Bleomycin in PBS RPMI Bleomycin in RPMI Ethylnitrosourea in RPMI
   Comet assay Specific	Detection method	Computerized image analysis system Manual analysis/categorization

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