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Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells

While tissue-resident monocytes and macrophages are considered to be vital players in the in vivo interaction between biomaterials and surrounding tissue, their isolation is limited. In order to establish in vitro models elucidating implant and tissue interactions, peripheral blood mononuclear cells...

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Autores principales: Klinder, Annett, Markhoff, Jana, Jonitz-Heincke, Anika, Sterna, Philipp, Salamon, Achim, Bader, Rainer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6395970/
https://www.ncbi.nlm.nih.gov/pubmed/30867690
http://dx.doi.org/10.3892/etm.2019.7204
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author Klinder, Annett
Markhoff, Jana
Jonitz-Heincke, Anika
Sterna, Philipp
Salamon, Achim
Bader, Rainer
author_facet Klinder, Annett
Markhoff, Jana
Jonitz-Heincke, Anika
Sterna, Philipp
Salamon, Achim
Bader, Rainer
author_sort Klinder, Annett
collection PubMed
description While tissue-resident monocytes and macrophages are considered to be vital players in the in vivo interaction between biomaterials and surrounding tissue, their isolation is limited. In order to establish in vitro models elucidating implant and tissue interactions, peripheral blood mononuclear cells (PBMCs) represent a viable source for bone marrow-derived monocytes and an alternative to tissue-resident cells. The aim of present study was to analyse different adhesion-preventing tissue culture plates for their potential to facilitate the culture of monocytes without differentiation into macrophages. Freshly isolated PBMCs were seeded into four commercially available tissue culture plates with different adhesive properties and were tested for surface CD14 and CD68 expression using flow cytometry following 7 days in culture. When PBMCs were cultivated in RPMI on Cellstar(®) Cell culture plates with Cell-Repellent Surface, a significant increase in CD14-positive cells was observed compared with cultivation in standard tissue culture-treated plates. This was accompanied by elevated cytokine production of interleukin-6 (IL6) and interleukin-8 (IL8); however, overall cell growth was not affected. When PBMCs were pre-cultured in cell-repellent plates, there was a higher yield of adherent cells after subsequent transfer into standard tissue culture-treated plates. Cultivation of PBMCs on cell-repellent culture plates favoured a monocytic phenotype and thus, represents an alternative to increase the fraction of monocytes yielded from PBMCs.
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spelling pubmed-63959702019-03-13 Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells Klinder, Annett Markhoff, Jana Jonitz-Heincke, Anika Sterna, Philipp Salamon, Achim Bader, Rainer Exp Ther Med Articles While tissue-resident monocytes and macrophages are considered to be vital players in the in vivo interaction between biomaterials and surrounding tissue, their isolation is limited. In order to establish in vitro models elucidating implant and tissue interactions, peripheral blood mononuclear cells (PBMCs) represent a viable source for bone marrow-derived monocytes and an alternative to tissue-resident cells. The aim of present study was to analyse different adhesion-preventing tissue culture plates for their potential to facilitate the culture of monocytes without differentiation into macrophages. Freshly isolated PBMCs were seeded into four commercially available tissue culture plates with different adhesive properties and were tested for surface CD14 and CD68 expression using flow cytometry following 7 days in culture. When PBMCs were cultivated in RPMI on Cellstar(®) Cell culture plates with Cell-Repellent Surface, a significant increase in CD14-positive cells was observed compared with cultivation in standard tissue culture-treated plates. This was accompanied by elevated cytokine production of interleukin-6 (IL6) and interleukin-8 (IL8); however, overall cell growth was not affected. When PBMCs were pre-cultured in cell-repellent plates, there was a higher yield of adherent cells after subsequent transfer into standard tissue culture-treated plates. Cultivation of PBMCs on cell-repellent culture plates favoured a monocytic phenotype and thus, represents an alternative to increase the fraction of monocytes yielded from PBMCs. D.A. Spandidos 2019-03 2019-01-25 /pmc/articles/PMC6395970/ /pubmed/30867690 http://dx.doi.org/10.3892/etm.2019.7204 Text en Copyright: © Klinder et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Klinder, Annett
Markhoff, Jana
Jonitz-Heincke, Anika
Sterna, Philipp
Salamon, Achim
Bader, Rainer
Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title_full Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title_fullStr Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title_full_unstemmed Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title_short Comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
title_sort comparison of different cell culture plates for the enrichment of non-adherent human mononuclear cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6395970/
https://www.ncbi.nlm.nih.gov/pubmed/30867690
http://dx.doi.org/10.3892/etm.2019.7204
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