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Open AccessArticle

Methods of Granulocyte Isolation from Human Blood and Labeling with Multimodal Superparamagnetic Iron Oxide Nanoparticles

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Hospital Israelita Albert Einstein, São Paulo 05652-900, Brazil
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LIM44—Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de São Paulo, São Paulo 01246-903, Brazil
*
Author to whom correspondence should be addressed.
Academic Editor: Nadine Millot
Molecules 2020, 25(4), 765; https://doi.org/10.3390/molecules25040765 (registering DOI)
Received: 11 December 2019 / Revised: 17 January 2020 / Accepted: 2 February 2020 / Published: 11 February 2020
(This article belongs to the Special Issue Advances in Metal Oxide Nanoparticles)
This in vitro study aimed to find the best method of granulocyte isolation for subsequent labeling with multimodal nanoparticles (magnetic and fluorescent properties) to enable detection by optical and magnetic resonance imaging (MRI) techniques. The granulocytes were obtained from venous blood samples from 12 healthy volunteers. To achieve high purity and yield, four different methods of granulocyte isolation were evaluated. The isolated granulocytes were labeled with multimodal superparamagnetic iron oxide nanoparticles (M-SPIONs) coated with dextran, and the iron load was evaluated qualitatively and quantitatively by MRI, near-infrared fluorescence (NIRF) and inductively coupled plasma mass spectrometry (ICP-MS). The best method of granulocyte isolation was Percoll with Ficoll, which showed 95.92% purity and 94% viability. After labeling with M-SPIONs, the granulocytes showed 98.0% purity with a yield of 3.5 × 106 cells/mL and more than 98.6% viability. The iron-loading value in the labeled granulocytes, as obtained by MRI, was 6.40 ± 0.18 pg/cell. Similar values were found with the ICP-MS and NIRF imaging techniques. Therefore, our study shows that it is possible to isolate granulocytes with high purity and yield and labeling with M-SPIONs provides a high internalized iron load and low toxicity to cells. Therefore, these M-SPION-labeled granulocytes could be a promising candidate for future use in inflammation/infection detection by optical and MRI techniques. View Full-Text
Keywords: granulocyte isolation; magnetic nanoparticle; SPION; magnetic resonance imaging; near-infrared fluorescence; ICP-MS; molecular imaging; inflammation; infection granulocyte isolation; magnetic nanoparticle; SPION; magnetic resonance imaging; near-infrared fluorescence; ICP-MS; molecular imaging; inflammation; infection
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Alvieri, F.; Mamani, J.B.; Nucci, M.P.; Oliveira, F.A.; Filgueiras, I.S.; Rego, G.N.A.; de Barboza, M.F.; da Silva, H.R.; Gamarra, L.F. Methods of Granulocyte Isolation from Human Blood and Labeling with Multimodal Superparamagnetic Iron Oxide Nanoparticles. Molecules 2020, 25, 765.

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