3rd International Conferences and Workshops on Basic and Applied
Sciences 2010 ISBN: 978-979-19096-1-7
B005
Evaluation of Hyperthermia Effect on Cell Viability by a Simple
Crystal Violet Method of MDA-MB 231 Cell Line
E.Asita and H.Salehhuddin
Department of Biological Science, Faculty of Bioscience and
Bioengineering,
University of Technology Malaysia, Johor, Malaysia e-mail:
[email protected]
email: [email protected]
Abstract
Viability of cultured mammalian cells is evaluated by a variety
of techniques. In this study, a simple crystal violet staining
method was used to determine hyperthermia effect on viability of
human breast cancer MDA-MB 231 cell. The cells were exposed to heat
at 42C for 2 hours to estimate the percentage of cell viability
using crystal violet procedure (4% paraformaldehyde fixation). The
value assay was measured continuously from day 1 to day 7. The
percentage of viability was significantly decreased after 7 days of
heat exposure at 42C (74.111.11%) compared to control (100%
viability) (p< 0.05). More cells were killed at high temperature
(42C) and longer duration of heat. However, temperatures above 42C
and longer exposure of heat (> 2 hours) can cause damages in
healthy cells. This study revealed that crystal violet was a
sensitive assay for adherent cell. It is also very effective;
simple; and rapid quantitative assay to measure the cell viability
of MDA-MB 231 cell. Keywords: Cell viability; Crystal violet assay;
Hyperthermia; Sensitive assay.
1 Introduction
A variety of methods have been used for evaluating the cytotoxic
effects of cultured cells, depending on the cell or tissue type.
The assays widely used in mammalian cell are based either on
colorimetric method or microscopic analysis (Juan et al., 2003).
One of the techniques, crystal violet (CV) staining assay is a well
known quantitative colorimetric procedure. The colorimetric
determination of the stained cells is based on loss of ability to
maintain and provide energy for metabolic growth and function
(Chiba et al., 1997). Crystal violet or gentian violet is a
triphenylmethane dye which taken up by cells (Jorge et al.,
2007). This technique has been used with modifications for a wide
number of applications, most of them to determine effect of
chemicals, drugs, or toxins from pathogens on cytotoxicity or cell
death (Harhaji et al., 2004; Shaik et al., 2004; Rothman, 1986 (tdk
ad dlm dapus)), and to evaluate cell viability (Thomas et al.,
2004) or proliferation of cell (Zivadinovc et al., 2005) under
different conditions.
The most common method used with adherent cells has been the
crystal violet assay when compared to other procedure such as
trypan blue method which the cells have to be physically or
enzymatically removed from the growth surface before measure the
cell viability (Sanford et al., 1951). Therefore, crystal violet
staining technique is very effective because it is simple;
reproducible assay of cytotoxicity; and a number of materials can
be evaluated simultaneously (Itagaki et al., 1991; Saotome et al.,
1989). Hyperthermia means that body tissue is exposed to high
temperatures (up to 45C). It is a type of cancer treatment (Samia
et al., 2007). Research has shown that high temperatures can damage
or kill cancer cells with usually minimal injury to normal tissue
(Van der zee, 2002). In the present study, it was aimed to
determine the effect of hyperthermia on viability in human breast
cancer MDA-MB 231 cell.
2 Methodology
2.1 Cells The human breast cancer cell line MDA-MB 231 was
obtained from American Type Culture Collection (ATCC) and cultured
in Dulbecco's modified Eagle's medium supplemented with 4.5 g/L
glucose, 1.5 g/L sodium bicarbonate, 100 U/mL penicillin, 100 mg/mL
streptomycin, and 10% fetal
3rd International Conferences and Workshops on Basic and Applied
Sciences 2010 ISBN: 978-979-19096-1-7
B005
reader at absorbance 595nm. This method has been widely used for
monitoring mammalian cell cultures because of the fast and
quantitative release of stained nuclei. Percentage viability of
MDA-MB 231 cell line determined by the crystal violet method; was
significantly and consistently high (p< 0.05). Therefore, it
proved that crystal violet was a sensitive assay for adherent cell.
It is also very effective; simple; and permits many samples to be
analyzed rapidly and simultaneously (Itagaki et al., 1991; Saotome
et al., 1989).
4 Conclusion
In this study, it revealed that crystal violet was a sensitive
and useful assay for adherent cell. Besides that, hyperthermia
enhanced reduction in viability of MDA-MB-231 cell.
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