RESEARCH PAPER
Effect of exopolysaccharide from Ganoderma applanatum on the electrical properties of mouse fibroblast cells line L929 culture using an electric cell-substrate impedance sensing (ECIS) – Preliminary study
More details
Hide details
1
Physiology Department, Medical University of Lublin, Poland
2
Department Biochemistry, Maria Curie-Skłodowska University, Lublin, Poland
3
ibidi GmbH Am Klopferspitz 19 D-82152 Martinsried (München) Germany
4
Institute of Electronics and Information Technology Technical University of Lublin, Poland
Ann Agric Environ Med. 2016;23(2):280-284
KEYWORDS
ABSTRACT
Introduction:
In recent years there has been intensified research on medicinal preparations of fungal origin. Some fungal polysaccharides may directly affect the inhibition of cancer cells proliferation which, stopping the cell cycle, leads to apoptosis. One of these substances (component of extract of Ganoderma spp) is extensively tested for its anti-cancer properties on the tumor cell lines. Electric cell-substrate impedance sensing (ECIS) is an in vitro impedance measuring system using alternating current (AC) to determinate the behaviour of the cells in physiological conditions.
Objective:
The aim of the study was to examine the electric properties (resistance, capacitance and impedance) of mouse fibroblasts cell line L929 after treatment by different concentration of crude exopolysaccharides from Ganoderma applanatum (GpEPS) in real time by ECIS technique.
Material and Methods:
For the study, the L929 cell line culture was treated by different concentrations of GpEPS: C1=228.5 µg/mL; C2=22.85 µg/mL; C3=2.285 µg/mL; C4=0.2285 µg/mL; and C5=0.02285 µg/mL. Default optimal frequencies were used: Resistance (R) 4000Hz, Impedance (Z) 16000Hz, Capacitance (C) 64000Hz.
Results:
The study demonstrated that GpEPS had no significant effect on the resistance, capacitance and impedance cells cultures, which implies that there is no significant effect on the physiological processes of L929 fibroblasts. This indicates the possibility of using GpEPS preparation in anti-cancer therapy.
Conclusions:
In the future, following further studies (comprising in preventive and therapeutic actions), GpEPS can be safely used in anti-cancer therapy which does not cause side-effects or damage to healthy cells.
REFERENCES (17)
1.
Paterson R R M. Ganoderma—a therapeutic fungal biofactory. Phytochemistry. 2006; 67(18): 1985–2001.
2.
Methacanon P, Madla S, Kirtikara K, Prasitsil M. Structural elucidation of bioactive fungi-derived polymers. Carbohydr Polym. 2005; 60(2): 199–203.
3.
Giaever I, Keese CR. Monitoring fibroblast behavior in tissue culture with an applied electric field . Proc. Natl Acad Sci. (USA) 1984; 81: 3761–3764.
4.
Giaever I, Keese CR. A morphological biosensor for mammalian cells. Nature, 1993, 366 (6455):591–592.
5.
Szulcek R, Bogaard H J, van Nieuw Amerongen G P. Electric Cell-substrate Impedance Sensing for the Quantification of Endothelial Proliferation, Barrier Function, and Motility. J Vis Exp. 2014; 85: e51300, doi:10.3791/51300.
6.
Xiao C, Lachance B, Sunahara G, and Luong J H T. Assessment of Cytotoxicity Using Electric Cell-Substrate Impedance Sensing: Concentration and Time Response Function Approach. Anal Chem. 2002; 74: 5748 – 5753.
7.
Osińska-Jaroszuk M, Jaszek M, Mizerska-Dudka M, Błachowicz A, Rejczak TP, Janusz G, et al. Exopolysaccharide from Ganoderma applanatum as a Promising Bioactive Compound with Cytostatic and Antibacterial Properties. Hindawi Publishing Corporation BioMed Res Int. 2014; Article ID 743812, 10 pages
http://dx.doi.org/10.1155/2014....
8.
Dubois M, Gilles K A, Hamilton J K, Rebers P A, and Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem. 1956; 28(3): 350–356.
9.
Hope C F A, Burns R G. Activity, origins and location of cellulases in a silt loam soil. Biol Fertil Soils. 1987; 5(2): 164–170.
10.
Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry. 1976; 72(1–2): 248–254.
11.
Malarczyk E. Transformation of phenolic acids by Nocardia. Acta Microbiol Polonica, 1998; 38(1): 45–53.
12.
Usui T, Iwasaki Y, Mizuno T, Tanaka M, Shinkai K, Arakawa M. Isolation and characterization of antitumor active b-d-glucans from the fruit bodies of Ganoderma applanatum. Carbohydrate Res. 1983; 115(1): 273–280.
13.
Lee WY, Park Y, Ahn JK, Ka K H, Park SY. Factors influencing the production of endopolysaccharide and exopolysaccharide from Ganoderma applanatum. Enzyme Microb Technol. 2007; 40(2): 249–254.
14.
Wegener J, Keese CR, Giaever I. Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. Exp Cell Res. 2000; 259(1): 158–166.
15.
Xiaoqiu H, Nguyen D, Greve DW, Domach MM. Simulation of microelectrode impedance changes due to cell growth. IEEE Sens J. 2004; 4(5): 576–583.
16.
Coffman FD and Cohen S. Impedance measurements in the biomedical Sciences. Anal Cellular Pathol. 2012; 35: 363–374 doi 10.3233/ACP-2012–0070.
17.
Ramasamy S, Bennet D, Kim S. Drug and bioactive molecule screening based on a bioelectrical impedance cell culture platform. Int J Nanomed. 2014; 9: 5789–5809.