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

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...

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Detalles Bibliográficos
Autores principales: Brás, Susana, França, Ângela, Cerca, Nuno
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301895/
https://www.ncbi.nlm.nih.gov/pubmed/32587796
http://dx.doi.org/10.7717/peerj.9295
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author Brás, Susana
França, Ângela
Cerca, Nuno
author_facet Brás, Susana
França, Ângela
Cerca, Nuno
author_sort Brás, Susana
collection PubMed
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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spelling pubmed-73018952020-06-24 Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes Brás, Susana França, Ângela Cerca, Nuno PeerJ Microbiology 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. PeerJ Inc. 2020-06-15 /pmc/articles/PMC7301895/ /pubmed/32587796 http://dx.doi.org/10.7717/peerj.9295 Text en ©2020 Brás et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Microbiology
Brás, Susana
França, Ângela
Cerca, Nuno
Optimizing a reliable ex vivo human blood model to analyze expression of Staphylococcus epidermidis genes
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 Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301895/
https://www.ncbi.nlm.nih.gov/pubmed/32587796
http://dx.doi.org/10.7717/peerj.9295
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