Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System

It is very important to assess cell viability rapidly and sensitively for the cell biology research, medical and pharmaceutical application. Compared to conventional methods, we have established a new rapid and sensitive bio-electrochemical system using small screen-printed carbon electrode (SPCE) a...

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Autores principales:	Mohammad Abdul ALIM, Minoru SUGA, Hiroaki SHINOHARA
Formato:	article
Lenguaje:	EN JA
Publicado:	The Electrochemical Society of Japan 2021
Materias:	electrochemical monitoring intracellular nadh cell viability double mediators Technology T Physical and theoretical chemistry QD450-801
Acceso en línea:	https://doaj.org/article/758c141660ab44288d17189d11f3beb1
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spelling	oai:doaj.org-article:758c141660ab44288d17189d11f3beb12021-11-05T00:17:27ZRapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System2186-245110.5796/electrochemistry.21-00088https://doaj.org/article/758c141660ab44288d17189d11f3beb12021-11-01T00:00:00Zhttps://www.jstage.jst.go.jp/article/electrochemistry/89/6/89_21-00088/_html/-char/enhttps://doaj.org/toc/2186-2451It is very important to assess cell viability rapidly and sensitively for the cell biology research, medical and pharmaceutical application. Compared to conventional methods, we have established a new rapid and sensitive bio-electrochemical system using small screen-printed carbon electrode (SPCE) and 1-methoxy-5-methylphenazinium methyl sulfate (mPMS)/[Fe(CN)6]3−(FeCN) as double electron mediators for monitoring cell viability through the measurement of intracellular NADH. A combination of 10 µmol L−1 (µM) mPMS and 500 µM FeCN was the optimum concentration and, 10 minutes (min) incubation was enough to monitor intracellular NADH by chronoamperometry at +0.5 V applications. This mPMS/FeCN system works as useful as previously reported menadione (Mena)/FeCN system. We confirmed that the electron transfer from intracellular NADH to mPMS occurred non-enzymatically, though the electron transfer from intracellular NADH to Mena was catalyzed by cytosolic enzyme. We applied our system to count the three kinds of mammalian cells. The oxidation current in chronoamperometry after 10 min incubation showed a good linear relationship in two times wider of cell concentration as compared to the cell concentration detected with water soluble tetrazolium-1 (WST-1) assay. The results indicated that the metabolically active mammalian cells could be quickly quantified by our method. Furthermore, we have applied this method to evaluate the acute cytotoxicity of oxamic acid on cytoma cells. Only 10 min incubation and high sensitivity embellished this method. These results strongly supported that our electrochemical method might be potent to alternate to WST assay for cell viability and acute cytotoxicity test.Mohammad Abdul ALIMMinoru SUGAHiroaki SHINOHARAThe Electrochemical Society of Japanarticleelectrochemical monitoringintracellular nadhcell viabilitydouble mediatorsTechnologyTPhysical and theoretical chemistryQD450-801ENJAElectrochemistry, Vol 89, Iss 6, Pp 579-584 (2021)
institution	DOAJ
collection	DOAJ
language	EN JA
topic	electrochemical monitoring intracellular nadh cell viability double mediators Technology T Physical and theoretical chemistry QD450-801
spellingShingle	electrochemical monitoring intracellular nadh cell viability double mediators Technology T Physical and theoretical chemistry QD450-801 Mohammad Abdul ALIM Minoru SUGA Hiroaki SHINOHARA Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
description	It is very important to assess cell viability rapidly and sensitively for the cell biology research, medical and pharmaceutical application. Compared to conventional methods, we have established a new rapid and sensitive bio-electrochemical system using small screen-printed carbon electrode (SPCE) and 1-methoxy-5-methylphenazinium methyl sulfate (mPMS)/[Fe(CN)6]3−(FeCN) as double electron mediators for monitoring cell viability through the measurement of intracellular NADH. A combination of 10 µmol L−1 (µM) mPMS and 500 µM FeCN was the optimum concentration and, 10 minutes (min) incubation was enough to monitor intracellular NADH by chronoamperometry at +0.5 V applications. This mPMS/FeCN system works as useful as previously reported menadione (Mena)/FeCN system. We confirmed that the electron transfer from intracellular NADH to mPMS occurred non-enzymatically, though the electron transfer from intracellular NADH to Mena was catalyzed by cytosolic enzyme. We applied our system to count the three kinds of mammalian cells. The oxidation current in chronoamperometry after 10 min incubation showed a good linear relationship in two times wider of cell concentration as compared to the cell concentration detected with water soluble tetrazolium-1 (WST-1) assay. The results indicated that the metabolically active mammalian cells could be quickly quantified by our method. Furthermore, we have applied this method to evaluate the acute cytotoxicity of oxamic acid on cytoma cells. Only 10 min incubation and high sensitivity embellished this method. These results strongly supported that our electrochemical method might be potent to alternate to WST assay for cell viability and acute cytotoxicity test.
format	article
author	Mohammad Abdul ALIM Minoru SUGA Hiroaki SHINOHARA
author_facet	Mohammad Abdul ALIM Minoru SUGA Hiroaki SHINOHARA
author_sort	Mohammad Abdul ALIM
title	Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
title_short	Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
title_full	Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
title_fullStr	Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
title_full_unstemmed	Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System
title_sort	rapid and highly sensitive electrochemical technique for cell viability assay via monitoring of intracellular nadh with new double mediator system
publisher	The Electrochemical Society of Japan
publishDate	2021
url	https://doaj.org/article/758c141660ab44288d17189d11f3beb1
work_keys_str_mv	AT mohammadabdulalim rapidandhighlysensitiveelectrochemicaltechniqueforcellviabilityassayviamonitoringofintracellularnadhwithnewdoublemediatorsystem AT minorusuga rapidandhighlysensitiveelectrochemicaltechniqueforcellviabilityassayviamonitoringofintracellularnadhwithnewdoublemediatorsystem AT hiroakishinohara rapidandhighlysensitiveelectrochemicaltechniqueforcellviabilityassayviamonitoringofintracellularnadhwithnewdoublemediatorsystem
_version_	1718444534780657664

Rapid and Highly Sensitive Electrochemical Technique for Cell Viability Assay via Monitoring of Intracellular NADH with New Double Mediator System

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