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Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes

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Resumo:Human blood is often used as an ex vivo model to mimic the environment encountered by pathogens inside the host. A significant variety of experimental conditions has been reported. However, optimization strategies are often not described. This study aimed to evaluate key parameters that are expected to influence Staphylococcus epidermidis gene expression when using human blood ex vivo models. Our data confirmed that blood antimicrobial activity was dependent on initial bacterial concentration. Furthermore, blood degradation over time resulted in lower antimicrobial activity, with a 2% loss of leukocytes viability correlating with a 5-fold loss of antimicrobial activity against S. epidermidis. We further demonstrated that the volume of human blood could be reduced to as little as 0.18 mL without affecting the stability of gene expression of the tested genes. Overall, the data described herein highlight experimental parameters that should be considered when using a human blood ex vivo model for S. epidermidis gene expression analysis.
Autores principais:Brás, Susana Maria Pereira
Outros Autores:França, Ângela Maria Oliveira Sousa; Cerca, Nuno
Assunto:Bacterial survival Staphylococcus epidermidis Human blood ex vivo model Volume of human blood Gene expression
Ano:2020
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Universidade do Minho
Idioma:inglês
Origem:RepositóriUM - Universidade do Minho
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author Brás, Susana Maria Pereira
author2 França, Ângela Maria Oliveira Sousa
Cerca, Nuno
author2_role author
author
author_facet Brás, Susana Maria Pereira
França, Ângela Maria Oliveira Sousa
Cerca, Nuno
author_role author
contributor_name_str_mv RepositóriUM - Universidade do Minho
country_str PT
creators_json_txt [{\"Person.name\":\"Brás, Susana Maria Pereira\"},{\"Person.name\":\"França, Ângela Maria Oliveira Sousa\"},{\"Person.name\":\"Cerca, Nuno\"}]
datacite.contributors.contributor.contributorName.fl_str_mv RepositóriUM - Universidade do Minho
datacite.creators.creator.creatorName.fl_str_mv Brás, Susana Maria Pereira
França, Ângela Maria Oliveira Sousa
Cerca, Nuno
datacite.date.Accepted.fl_str_mv 2020-06-15T00:00:00Z
datacite.date.available.fl_str_mv 2020-06-23T13:10:19Z
datacite.date.embargoed.fl_str_mv 2020-06-23T13:10:19Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.subjects.subject.fl_str_mv Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
datacite.titles.title.fl_str_mv Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
dc.contributor.none.fl_str_mv RepositóriUM - Universidade do Minho
dc.creator.none.fl_str_mv Brás, Susana Maria Pereira
França, Ângela Maria Oliveira Sousa
Cerca, Nuno
dc.date.Accepted.fl_str_mv 2020-06-15T00:00:00Z
dc.date.available.fl_str_mv 2020-06-23T13:10:19Z
dc.date.embargoed.fl_str_mv 2020-06-23T13:10:19Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv https://hdl.handle.net/1822/65740
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv Peerj, Inc.
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
dc.title.fl_str_mv Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description Human blood is often used as an ex vivo model to mimic the environment encountered by pathogens inside the host. A significant variety of experimental conditions has been reported. However, optimization strategies are often not described. This study aimed to evaluate key parameters that are expected to influence Staphylococcus epidermidis gene expression when using human blood ex vivo models. Our data confirmed that blood antimicrobial activity was dependent on initial bacterial concentration. Furthermore, blood degradation over time resulted in lower antimicrobial activity, with a 2% loss of leukocytes viability correlating with a 5-fold loss of antimicrobial activity against S. epidermidis. We further demonstrated that the volume of human blood could be reduced to as little as 0.18 mL without affecting the stability of gene expression of the tested genes. Overall, the data described herein highlight experimental parameters that should be considered when using a human blood ex vivo model for S. epidermidis gene expression analysis.
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eu_rights_str_mv openAccess
format article
fulltext.url.fl_str_mv https://repositorium.uminho.pt/bitstreams/5c25cd47-6d59-4d31-95fb-fcf77d16c325/download
id rum_4db54ea5d4d4deaef8ddbf7278652ed0
identifier.url.fl_str_mv https://hdl.handle.net/1822/65740
instacron_str repositorium
institution Universidade do Minho
instname_str Universidade do Minho
language eng
network_acronym_str rum
network_name_str RepositóriUM - Universidade do Minho
oai_identifier_str oai:repositorium.uminho.pt:1822/65740
organization_str_mv urn:organizationAcronym:repositorium
person_str_mv Brás, Susana Maria Pereira
França, Ângela Maria Oliveira Sousa
Cerca, Nuno
publishDate 2020
publisher.none.fl_str_mv Peerj, Inc.
reponame_str RepositóriUM - Universidade do Minho
repository_id_str urn:repositoryAcronym:rum
service_str_mv urn:repositoryAcronym:rum
spelling engPeerj, Inc.porHuman blood is often used as an ex vivo model to mimic the environment encountered by pathogens inside the host. A significant variety of experimental conditions has been reported. However, optimization strategies are often not described. This study aimed to evaluate key parameters that are expected to influence Staphylococcus epidermidis gene expression when using human blood ex vivo models. Our data confirmed that blood antimicrobial activity was dependent on initial bacterial concentration. Furthermore, blood degradation over time resulted in lower antimicrobial activity, with a 2% loss of leukocytes viability correlating with a 5-fold loss of antimicrobial activity against S. epidermidis. We further demonstrated that the volume of human blood could be reduced to as little as 0.18 mL without affecting the stability of gene expression of the tested genes. Overall, the data described herein highlight experimental parameters that should be considered when using a human blood ex vivo model for S. epidermidis gene expression analysis.application/pdfporOptimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genesBrás, Susana Maria PereiraFrança, Ângela Maria Oliveira SousaCerca, NunoHostingInstitutionOrganizationalRepositóriUM - Universidade do Minhoe-mailmailto:repositorium@usdb.uminho.ptrepositorium@usdb.uminho.ptCITATIONBrás, Susana; França, Angela; Cerca, Nuno, Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes. PeerJ, 8(e9295), 2020ISSNIsPartOf2167-8359DOIIsPartOf10.7717/peerj.92952020-06-23T13:10:19Z2020-06-152020-06-20T09:59:28Z2020-06-15T00:00:00ZHandlehttps://hdl.handle.net/1822/65740http://purl.org/coar/access_right/c_abf2open accessBacterial survivalStaphylococcus epidermidisHuman blood ex vivo modelVolume of human bloodGene expression490806 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://repositorium.uminho.pt/bitstreams/5c25cd47-6d59-4d31-95fb-fcf77d16c325/download
spellingShingle Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
Brás, Susana Maria Pereira
Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
status SINGLETON
subject.fl_str_mv Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
title Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
title_full Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
title_fullStr Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
title_full_unstemmed Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
title_short Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
title_sort Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
topic Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
topic_facet Bacterial survival
Staphylococcus epidermidis
Human blood ex vivo model
Volume of human blood
Gene expression
url https://hdl.handle.net/1822/65740
visible 1