Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells

Exposure to asbestos fibers increases the risk of mesothelioma in humans. One hypothetical carcinogenic mechanism is that asbestos fibers may directly induce mutations in mesothelial cells. Although the uptake of asbestos fibers by mesothelial cells is recognized, methods for the quantification of t...

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Main Authors:	Yamashita, Kyoko, Nagai, Hirotaka, Kondo, Yuji, Misawa, Nobuaki, Toyokuni, Shinya
Format:	Online
Language:	English
Published:	the Society for Free Radical Research Japan 2013
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3705149/

id	pubmed-3705149
recordtype	oai_dc
spelling	pubmed-37051492013-07-19 Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells Yamashita, Kyoko Nagai, Hirotaka Kondo, Yuji Misawa, Nobuaki Toyokuni, Shinya Original Article Exposure to asbestos fibers increases the risk of mesothelioma in humans. One hypothetical carcinogenic mechanism is that asbestos fibers may directly induce mutations in mesothelial cells. Although the uptake of asbestos fibers by mesothelial cells is recognized, methods for the quantification of the uptake have not been well established. In the present study, we evaluated two distinct methods, using crocidolite fibers and MeT5A mesothelial cells. One method is histological evaluation using the cell-block technique, which allows for the direct cross-sectional observation of cells and fibers. We found the bright field observation with ×1000 magnification (oil-immersion) of the sample with Kernechtrot staining was most suitable for this purpose. The other method is flow cytometric analysis, which permits the evaluation of a much larger number of cells. We observed that the side scatter (SSC) increased with the intracellular fibers, and that the “mean SSC ratio (treated/control)” was useful for quantification. We could collect the cells with abundant internalized crocidolite fibers by sorting. Results of the two methodologies were correlated well in the experiments. The quantities of internalized fibers increased with incubation time and loaded dosage, but they were inversely associated with cellular density in culture. the Society for Free Radical Research Japan 2013-07 2013-06-01 /pmc/articles/PMC3705149/ /pubmed/23874067 http://dx.doi.org/10.3164/jcbn.12-104 Text en Copyright © 2013 JCBN This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Yamashita, Kyoko Nagai, Hirotaka Kondo, Yuji Misawa, Nobuaki Toyokuni, Shinya
spellingShingle	Yamashita, Kyoko Nagai, Hirotaka Kondo, Yuji Misawa, Nobuaki Toyokuni, Shinya Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
author_facet	Yamashita, Kyoko Nagai, Hirotaka Kondo, Yuji Misawa, Nobuaki Toyokuni, Shinya
author_sort	Yamashita, Kyoko
title	Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
title_short	Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
title_full	Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
title_fullStr	Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
title_full_unstemmed	Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
title_sort	evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells
description	Exposure to asbestos fibers increases the risk of mesothelioma in humans. One hypothetical carcinogenic mechanism is that asbestos fibers may directly induce mutations in mesothelial cells. Although the uptake of asbestos fibers by mesothelial cells is recognized, methods for the quantification of the uptake have not been well established. In the present study, we evaluated two distinct methods, using crocidolite fibers and MeT5A mesothelial cells. One method is histological evaluation using the cell-block technique, which allows for the direct cross-sectional observation of cells and fibers. We found the bright field observation with ×1000 magnification (oil-immersion) of the sample with Kernechtrot staining was most suitable for this purpose. The other method is flow cytometric analysis, which permits the evaluation of a much larger number of cells. We observed that the side scatter (SSC) increased with the intracellular fibers, and that the “mean SSC ratio (treated/control)” was useful for quantification. We could collect the cells with abundant internalized crocidolite fibers by sorting. Results of the two methodologies were correlated well in the experiments. The quantities of internalized fibers increased with incubation time and loaded dosage, but they were inversely associated with cellular density in culture.
publisher	the Society for Free Radical Research Japan
publishDate	2013
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3705149/
_version_	1611993017998114816

Evaluation of two distinct methods to quantify the uptake of crocidolite fibers by mesothelial cells

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