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Article: Modification of the MTT method for the study of bilirubin cytotoxicity

TitleModification of the MTT method for the study of bilirubin cytotoxicity
Authors
KeywordsBilirubin
Cell viability assay
Cytotoxicity
Mitochondrial function assay
Modified MTT method
Issue Date1998
Citation
Acta Paediatrica Japonica (Overseas Edition), 1998, v. 40 n. 4, p. 313-317 How to Cite?
AbstractBackground: We propose a modification of the MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method to study the cytotoxicity of bilirubin. The original method involves reading the intensity of a purplish blue color resulting from the conversion of MTT to formazan crystals by the mitochondria of viable cells. We have found that when the method is applied to study the effect of bilirubin on growing cells, precipitation of the yellow bilirubin pigment interferes with the colorimetric reading. Methods: A human liver cell line was used. The interference of bilirubin deposition on the MTT assay was investigated by comparing the value of optical density of the MTT solution in the presence and absence of bilirubin. The effect of 0.04 mol/L HCL-isopropanol on the bilirubin precipitate was tested by recovering the amount of bilirubin from the wells after the isopropanol treatment. Results: Bilirubin deposition increases MTT reading by 10-24%. Hydrochloride-isopropanol (0.04 mol/L) dissolves MTT formazan only without disturbing the bilirubin precipitates. The bilirubin extracted into the supernatant was less than 5% of the total bilirubin deposited. Discussion: This indirect MTT assay, as developed in this study, could eliminate the interference of bilirubin deposits and serve as a good method for the study of bilirubin cytotoxicity.
Persistent Identifierhttp://hdl.handle.net/10722/79882
ISSN
2000 Impact Factor: 0.523
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorNgai, KCen_HK
dc.contributor.authorYeung, CYen_HK
dc.contributor.authorKarlberg, Jen_HK
dc.date.accessioned2010-09-06T07:59:48Z-
dc.date.available2010-09-06T07:59:48Z-
dc.date.issued1998en_HK
dc.identifier.citationActa Paediatrica Japonica (Overseas Edition), 1998, v. 40 n. 4, p. 313-317en_HK
dc.identifier.issn0374-5600en_HK
dc.identifier.urihttp://hdl.handle.net/10722/79882-
dc.description.abstractBackground: We propose a modification of the MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method to study the cytotoxicity of bilirubin. The original method involves reading the intensity of a purplish blue color resulting from the conversion of MTT to formazan crystals by the mitochondria of viable cells. We have found that when the method is applied to study the effect of bilirubin on growing cells, precipitation of the yellow bilirubin pigment interferes with the colorimetric reading. Methods: A human liver cell line was used. The interference of bilirubin deposition on the MTT assay was investigated by comparing the value of optical density of the MTT solution in the presence and absence of bilirubin. The effect of 0.04 mol/L HCL-isopropanol on the bilirubin precipitate was tested by recovering the amount of bilirubin from the wells after the isopropanol treatment. Results: Bilirubin deposition increases MTT reading by 10-24%. Hydrochloride-isopropanol (0.04 mol/L) dissolves MTT formazan only without disturbing the bilirubin precipitates. The bilirubin extracted into the supernatant was less than 5% of the total bilirubin deposited. Discussion: This indirect MTT assay, as developed in this study, could eliminate the interference of bilirubin deposits and serve as a good method for the study of bilirubin cytotoxicity.en_HK
dc.languageengen_HK
dc.relation.ispartofActa Paediatrica Japonica (Overseas Edition)en_HK
dc.subjectBilirubinen_HK
dc.subjectCell viability assayen_HK
dc.subjectCytotoxicityen_HK
dc.subjectMitochondrial function assayen_HK
dc.subjectModified MTT methoden_HK
dc.titleModification of the MTT method for the study of bilirubin cytotoxicityen_HK
dc.typeArticleen_HK
dc.identifier.emailKarlberg, J: jpekarl@hkucc.hku.hken_HK
dc.identifier.authorityKarlberg, J=rp00400en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid9745771-
dc.identifier.scopuseid_2-s2.0-0031846693en_HK
dc.identifier.hkuros39390en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0031846693&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume40en_HK
dc.identifier.issue4en_HK
dc.identifier.spage313en_HK
dc.identifier.epage317en_HK
dc.identifier.isiWOS:000075572400004-
dc.identifier.scopusauthoridNgai, KC=7101628546en_HK
dc.identifier.scopusauthoridYeung, CY=7201354144en_HK
dc.identifier.scopusauthoridKarlberg, J=7005218406en_HK

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