Marine fish primary hepatocyte isolation and culture

Detalhes bibliográficos
Autor(a) principal: Figueiredo, Neusa
Data de Publicação: 2021
Outros Autores: Matos, Beatriz, Diniz, Mário Sousa, Branco, Vasco, Martins, Marta
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/133056
Resumo: PTDC/CTA-AMB/29173/2017 UIDB04292/2020 UIDP/04292/2020 UIDB04378/2020 UIDP/04378/2020 UIDB/04138/2020 UIDP/04138/ 2020 PTDC/CTA-AMB/29173/2017, NO-VAID39 grant 2020.09005. BD. DL57/2016/CP1376/CT002 CEECINST/00102/2018
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spelling Marine fish primary hepatocyte isolation and cultureNew insights to enzymatic dissociation pancreatin digestionIn vitro assaysPrimary hepatocytesSparus aurataPollutionPublic Health, Environmental and Occupational HealthHealth, Toxicology and MutagenesisSDG 3 - Good Health and Well-beingSDG 14 - Life Below WaterPTDC/CTA-AMB/29173/2017 UIDB04292/2020 UIDP/04292/2020 UIDB04378/2020 UIDP/04378/2020 UIDB/04138/2020 UIDP/04138/ 2020 PTDC/CTA-AMB/29173/2017, NO-VAID39 grant 2020.09005. BD. DL57/2016/CP1376/CT002 CEECINST/00102/2018Primary cell cultures from wild organisms have been gaining relevance in ecotoxicology as they are considered more sensitive than immortalized cell lines and retain the biochemical pathways found in vivo. In this study, the efficacy of two methods for primary hepatocyte cell isolation was compared using liver from two marine fish (Sparus aurata and Psetta maxima): (i) two-step collagenase perfusion and (ii) pancreatin digestion with modifications. Cell cultures were incubated in L-15 medium at 17 ± 1 °C and monitored for up to six days for cell viability and function using the trypan blue exclusion test, MTT test, lactate dehydrogenase (LDH) activity, and ethoxyresorufin O-deethylase (EROD) activity after Benzo[a]Pyrene exposure. The results showed significant differences between the number of viable cells (p < 0.05), the highest number being obtained for the pancreatin digestion method (average = 4.5 ± 1.9 × 107 cells). Moreover, the hepatocytes showed solid adherence to the culture plate and the rounded shape, changing into a triangular/polygonal shape. The cell viability and function obtained by pancreatin digestion were maintained for five days, and the EROD induction after exposure to the B[a]P showed that cells were metabolically active. This study shows that the optimized pancreatin digestion method is a valid, cost-effective, and simple alternative to the standard perfusion method for the isolation of primary hepatocytes from fish and is suitable for ecotoxicological studies involving marine pollutants, such as PAHs.MARE - Centro de Ciências do Mar e do AmbienteDCEA - Departamento de Ciências e Engenharia do AmbienteUCIBIO - Applied Molecular Biosciences UnitRUNFigueiredo, NeusaMatos, BeatrizDiniz, Mário SousaBranco, VascoMartins, Marta2022-02-16T23:19:22Z2021-02-032021-02-03T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article14application/pdfhttp://hdl.handle.net/10362/133056eng1661-7827PURE: 36673207https://doi.org/10.3390/ijerph18041380info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:11:44Zoai:run.unl.pt:10362/133056Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:47:41.075766Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Marine fish primary hepatocyte isolation and culture
New insights to enzymatic dissociation pancreatin digestion
title Marine fish primary hepatocyte isolation and culture
spellingShingle Marine fish primary hepatocyte isolation and culture
Figueiredo, Neusa
In vitro assays
Primary hepatocytes
Sparus aurata
Pollution
Public Health, Environmental and Occupational Health
Health, Toxicology and Mutagenesis
SDG 3 - Good Health and Well-being
SDG 14 - Life Below Water
title_short Marine fish primary hepatocyte isolation and culture
title_full Marine fish primary hepatocyte isolation and culture
title_fullStr Marine fish primary hepatocyte isolation and culture
title_full_unstemmed Marine fish primary hepatocyte isolation and culture
title_sort Marine fish primary hepatocyte isolation and culture
author Figueiredo, Neusa
author_facet Figueiredo, Neusa
Matos, Beatriz
Diniz, Mário Sousa
Branco, Vasco
Martins, Marta
author_role author
author2 Matos, Beatriz
Diniz, Mário Sousa
Branco, Vasco
Martins, Marta
author2_role author
author
author
author
dc.contributor.none.fl_str_mv MARE - Centro de Ciências do Mar e do Ambiente
DCEA - Departamento de Ciências e Engenharia do Ambiente
UCIBIO - Applied Molecular Biosciences Unit
RUN
dc.contributor.author.fl_str_mv Figueiredo, Neusa
Matos, Beatriz
Diniz, Mário Sousa
Branco, Vasco
Martins, Marta
dc.subject.por.fl_str_mv In vitro assays
Primary hepatocytes
Sparus aurata
Pollution
Public Health, Environmental and Occupational Health
Health, Toxicology and Mutagenesis
SDG 3 - Good Health and Well-being
SDG 14 - Life Below Water
topic In vitro assays
Primary hepatocytes
Sparus aurata
Pollution
Public Health, Environmental and Occupational Health
Health, Toxicology and Mutagenesis
SDG 3 - Good Health and Well-being
SDG 14 - Life Below Water
description PTDC/CTA-AMB/29173/2017 UIDB04292/2020 UIDP/04292/2020 UIDB04378/2020 UIDP/04378/2020 UIDB/04138/2020 UIDP/04138/ 2020 PTDC/CTA-AMB/29173/2017, NO-VAID39 grant 2020.09005. BD. DL57/2016/CP1376/CT002 CEECINST/00102/2018
publishDate 2021
dc.date.none.fl_str_mv 2021-02-03
2021-02-03T00:00:00Z
2022-02-16T23:19:22Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/133056
url http://hdl.handle.net/10362/133056
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1661-7827
PURE: 36673207
https://doi.org/10.3390/ijerph18041380
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 14
application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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