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Effects of sample collection and storage conditions on DNA damage in buccal cells from agricultural workers.

Authors
Muniz-JF; McCauley-LA; Pak-V; Lasarev-MR; Kisby-GE
Source
Mutat Res Genet Toxicol Environ Mutagen 2011 Feb; 720(1-2):8-13
NIOSHTIC No.
20038622
Abstract
Buccal cells are becoming a widely used tissue source for monitoring human exposure to occupational and environmental genotoxicants. A variety of methods exist for collecting buccal cells from the oral cavity, including rinsing with saline, mouthwash, or scraping the oral cavity. Buccal cells are also routinely cryopreserved with dimethyl sulfoxide (DMSO), then examined later for DNA damage by the comet assay. The effects of these different sampling procedures on the integrity of buccal cells for measuring DNA damage are unknown. This study examined the influence of the collection and cryopreservation of buccal cells on cell survival and DNA integrity. In individuals who rinsed with Hank's balanced salt solution (HBSS), the viability of leukocytes (90%) was significantly (p < 0.01) greater than that of epithelial cells (12%). Similar survival rates were found for leukocytes (88%) and epithelial cells (10%) after rinsing with Listerine® mouthwash. However, the viability of leukocytes after cryopreservation varied significantly (p < 0.01) with DMSO concentration. Cell survival was greatest at 5% DMSO. Cryopreservation also influenced the integrity of DNA in the comet assay. Although tail length and tail moment were comparable in fresh or cryopreserved samples, the average head intensity for cryopreserved samples was approximately 6 units lower (95% CI: 0.8-12 units lower) than for fresh samples (t25 = -2.36, p = 0.026). These studies suggest that the collection and storage of buccal samples are critical factors for the assessment of DNA damage. Moreover, leukocytes appear to be a more reliable source of human tissue for assessing DNA damage and possibly other biochemical changes.
Keywords
Biological-effects; Biological-monitoring; Cell-damage; Cell-transformation; Cellular-reactions; DNA-damage; Environmental-exposure; Farmers; Genotoxic-effects; Genotoxicity; Author Keywords: Comet assay; Leukocytes; Epithelial cells; DMSO; Cryopreservation
Contact
Juan F. Muniz, School of Nursing, University of Pennsylvania, 418 Curie Blvd., Philadelphia, PA 19104-6096
CODEN
MRGMFI
CAS No.
67-68-5
Publication Date
20110228
Document Type
Journal Article
Email Address
jfmuniz@nursing.upenn.edu
Funding Type
Grant
Fiscal Year
2011
NTIS Accession No.
NTIS Price
Identifying No.
Grant-Number-R01-OH-008057
Issue of Publication
1-2
ISSN
1383-5718
Source Name
Mutation Research - Genetic Toxicology and Environmental Mutagenesis
State
OR; PA
Performing Organization
University Of Pennsylvania
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