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Article: The OECD validation program of the H295R Steroidogenesis Assay for the identification of in vitro inhibitors and inducers of testosterone and estradiol production. Phase 2: Inter-laboratory pre-validation studies

TitleThe OECD validation program of the H295R Steroidogenesis Assay for the identification of in vitro inhibitors and inducers of testosterone and estradiol production. Phase 2: Inter-laboratory pre-validation studies
Authors
KeywordsEndocrine disruption
Estradiol
H295R
Hormone production
OECD
Sex steroid
Steroidogenesis
Testosterone
Transferability
Validation
Issue Date2007
PublisherSpringer. The Journal's web site is located at http://www.springer.com/environment/journal/11356
Citation
Environmental Science And Pollution Research, 2007, v. 14 n. 1 SPEC. ISS., p. 23-30 How to Cite?
AbstractBackground, Goals and Scope. In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the 'Special Activity on the Testing and Assessment of Endocrine Disruptors' within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design. Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results. All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC50s calculated were comparable (coefficients of variation 34-49%) for all hormones. Discussion and Perspectives. The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. © 2007 ecomed publishers (Verlagsgruppe Hüthig Jehle Rehm GmbH).
Persistent Identifierhttp://hdl.handle.net/10722/92740
ISSN
2015 Impact Factor: 2.76
2015 SCImago Journal Rankings: 0.886
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorHecker, Men_HK
dc.contributor.authorHollert, Hen_HK
dc.contributor.authorCooper, Ren_HK
dc.contributor.authorVinggaard, AMen_HK
dc.contributor.authorAkahori, Yen_HK
dc.contributor.authorMurphy, Men_HK
dc.contributor.authorNellemann, Cen_HK
dc.contributor.authorHigley, Een_HK
dc.contributor.authorNewsted, Jen_HK
dc.contributor.authorWu, Ren_HK
dc.contributor.authorLam, Pen_HK
dc.contributor.authorLaskey, Jen_HK
dc.contributor.authorBuckalew, Aen_HK
dc.contributor.authorGrund, Sen_HK
dc.contributor.authorNakai, Men_HK
dc.contributor.authorTimm, Gen_HK
dc.contributor.authorGiesy, Jen_HK
dc.date.accessioned2010-09-17T10:55:46Z-
dc.date.available2010-09-17T10:55:46Z-
dc.date.issued2007en_HK
dc.identifier.citationEnvironmental Science And Pollution Research, 2007, v. 14 n. 1 SPEC. ISS., p. 23-30en_HK
dc.identifier.issn0944-1344en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92740-
dc.description.abstractBackground, Goals and Scope. In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the 'Special Activity on the Testing and Assessment of Endocrine Disruptors' within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design. Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results. All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC50s calculated were comparable (coefficients of variation 34-49%) for all hormones. Discussion and Perspectives. The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. © 2007 ecomed publishers (Verlagsgruppe Hüthig Jehle Rehm GmbH).en_HK
dc.languageengen_HK
dc.publisherSpringer. The Journal's web site is located at http://www.springer.com/environment/journal/11356en_HK
dc.relation.ispartofEnvironmental Science and Pollution Researchen_HK
dc.subjectEndocrine disruptionen_HK
dc.subjectEstradiolen_HK
dc.subjectH295Ren_HK
dc.subjectHormone productionen_HK
dc.subjectOECDen_HK
dc.subjectSex steroiden_HK
dc.subjectSteroidogenesisen_HK
dc.subjectTestosteroneen_HK
dc.subjectTransferabilityen_HK
dc.subjectValidationen_HK
dc.titleThe OECD validation program of the H295R Steroidogenesis Assay for the identification of in vitro inhibitors and inducers of testosterone and estradiol production. Phase 2: Inter-laboratory pre-validation studiesen_HK
dc.typeArticleen_HK
dc.identifier.emailWu, R: rudolfwu@hku.hken_HK
dc.identifier.authorityWu, R=rp01398en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1065/espr2007.03.402en_HK
dc.identifier.scopuseid_2-s2.0-34248335900en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-34248335900&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume14en_HK
dc.identifier.issue1 SPEC. ISS.en_HK
dc.identifier.spage23en_HK
dc.identifier.epage30en_HK
dc.identifier.isiWOS:000246119700004-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridHecker, M=35247848500en_HK
dc.identifier.scopusauthoridHollert, H=6701325288en_HK
dc.identifier.scopusauthoridCooper, R=7404337959en_HK
dc.identifier.scopusauthoridVinggaard, AM=6701813758en_HK
dc.identifier.scopusauthoridAkahori, Y=16314791000en_HK
dc.identifier.scopusauthoridMurphy, M=7403900446en_HK
dc.identifier.scopusauthoridNellemann, C=7004128077en_HK
dc.identifier.scopusauthoridHigley, E=13004073100en_HK
dc.identifier.scopusauthoridNewsted, J=6603677236en_HK
dc.identifier.scopusauthoridWu, R=7402945079en_HK
dc.identifier.scopusauthoridLam, P=7202365776en_HK
dc.identifier.scopusauthoridLaskey, J=7003887713en_HK
dc.identifier.scopusauthoridBuckalew, A=6603278375en_HK
dc.identifier.scopusauthoridGrund, S=24780533100en_HK
dc.identifier.scopusauthoridNakai, M=7201803566en_HK
dc.identifier.scopusauthoridTimm, G=7006462733en_HK
dc.identifier.scopusauthoridGiesy, J=35459135300en_HK

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